TREM compositions and uses thereof

文档序号：1966736 发布日期：2021-12-14 浏览：16次中文

阅读说明：本技术 Trem组合物及其用途 (TREM compositions and uses thereof ) 是由 D.A.贝里 T.阿纳斯塔西亚迪斯 N.B.阿费扬 C.E.哈伊丁于 2020-01-17 设计创作，主要内容包括：本发明总体上涉及基于tRNA的效应子分子和与其相关的方法。(The invention relates generally to tRNA-based effector molecules and methods related thereto.)

1. A method of making a purified tRNA effector molecule (TREM) pharmaceutical composition, the method comprising:

providing a mammalian host cell comprising an exogenous nucleic acid, e.g., DNA or RNA, encoding the TREM;

maintaining the mammalian cell under conditions sufficient to express the TREM;

purifying the TREM from the mammalian host cell, e.g., according to the methods described herein; and

formulating the purified TREM into a pharmaceutical composition, e.g. by combining said TREM with a pharmaceutical excipient,

thereby preparing the TREM pharmaceutical composition.

2. The method of claim 1, wherein the nucleic acid comprises RNA that, upon reverse transcription, produces DNA that is transcribable into TREM.

3. The method of claim 1 or 2, wherein the nucleic acid comprises an RNA sequence that is at least 90% identical to an RNA sequence encoded by a DNA sequence listed in table 1, or a fragment or functional fragment thereof.

4. The method of claim 1 or 2, wherein the nucleic acid comprises an RNA sequence comprising a consensus sequence provided herein.

5. The method of any one of the preceding claims, wherein the mammalian host cell is selected from the group consisting of: non-human cells or cell lines, or human cells or cell lines, such as HEK293T cells (e.g., Freestyle 293-F cells), HT-1080 cells, PER. C6 cells, HKB-11 cells, CAP cells, HuH-7 cells, BHK 21 cells, MRC-S cells, MDCK cells, VERO cells, WI-38 cells, Chinese Hamster Ovary (CHO) cells, or MCF7 cells.

6. The method of any one of the preceding claims, wherein the purification step comprises one, two or all of the following steps, e.g., in the order:

(i) separating the nucleic acids from the cell debris to provide an RNA preparation;

(ii) separating RNA less than a threshold number of nucleotides, e.g., less than 500nt, less than 400nt, less than 300nt, less than 250nt, less than 200nt, less than 150nt, from larger RNA species in the RNA preparation to produce a small RNA preparation; and/or

(iii) TREM is separated from other RNA species in the small RNA preparation by affinity-based separation, e.g., sequence affinity-based separation.

7. A composition comprising a purified tRNA effector molecule (TREM) (e.g., a purified TREM composition prepared according to a method described herein), the purified tRNA effector molecule comprising:

(i) an RNA sequence at least 90% identical to an RNA sequence encoded by a DNA sequence listed in table 1, or a fragment or functional fragment thereof; or

(ii) RNA sequences comprising the consensus sequences provided herein.

8. A GMP-grade recombinant TREM composition (e.g., a TREM composition that conforms to cGMP and/or is prepared according to similar requirements), comprising:

(i) an RNA sequence at least 90% identical to an RNA sequence encoded by a DNA sequence listed in table 1, or a fragment or functional fragment thereof; or

(ii) RNA sequences comprising the consensus sequences provided herein.

9. The TREM composition of claim 7 or 8, wherein the composition comprises one or more, e.g., a plurality of TREMs.

10. The TREM composition of any one of claims 7 to 9, wherein the composition comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 species of TREM.

11. The TREM composition of any one of claims 7 to 10, wherein the TREM composition (or an intermediate in the production of a TREM composition) comprises one or more of the following characteristics:

(i) Purity of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%;

(ii) host Cell Protein (HCP) contamination of less than 0.1ng/ml, 1ng/ml, 5ng/ml, 10ng/ml, 15ng/ml, 20ng/ml, 25ng/ml, 30ng/ml, 35ng/ml, 40ng/ml, 50ng/ml, 60ng/ml, 70ng/ml, 80ng/ml, 90ng/ml or 100 ng/ml;

(iii) (ii) Host Cell Protein (HCP) contamination of less than 0.1ng, 1ng, 5ng, 10ng, 15ng, 20ng, 25ng, 30ng, 35ng, 40ng, 50ng, 60ng, 70ng, 80ng, 90ng, or 100ng per milligram (mg) of the TREM composition;

(iv) DNA, e.g., host cell DNA, of less than 1ng/ml, 5ng/ml, 10ng/ml, 15ng/ml, 20ng/ml, 25ng/ml, 30ng/ml, 35ng/ml, 40ng/ml, 50ng/ml, 60ng/ml, 70ng/ml, 80ng/ml, 90ng/ml or 100 ng/ml;

(v) less than 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% TREM fragments relative to full-length TREM;

(vi) low or absent endotoxin levels, e.g., a negative result as measured by a Limulus Amoebocyte Lysate (LAL) test;

(vii) in vitro translation activity, e.g., as measured by the assay described in example 15;

(viii) TREM concentration of at least 0.1ng/mL, 0.5ng/mL, 1ng/mL, 5ng/mL, 10ng/mL, 50ng/mL, 0.1ug/mL, 0.5ug/mL, 1ug/mL, 2ug/mL, 5ug/mL, 10ug/mL, 20ug/mL, 30ug/mL, 40ug/mL, 50ug/mL, 60ug/mL, 70ug/mL, 80ug/mL, 100ug/mL, 200ug/mL, 300ug/mL, 500ug/mL, 1000ug/mL, 5000ug/mL, 10,000ug/mL, or 100,000 ug/mL;

(ix) aseptically, e.g., according to the cGMP guidelines for sterile pharmaceutical products, e.g., the composition or formulation supports the growth of less than 100 viable microorganisms when tested under sterile conditions, the composition or formulation meets the criteria of USP <71>, and/or the composition or formulation meets the criteria of USP <85 >; or

(x) Viral contamination, e.g., the composition or formulation is not present or no level of viral contamination is detected.

12. A method of modulating a tRNA pool in a cell, the method comprising:

providing a purified TREM composition, and contacting the cell with the TREM composition,

thereby modulating the tRNA pool in the cell.

13. The method of claim 12, wherein the TREM composition is prepared by:

providing a mammalian host cell comprising an exogenous nucleic acid, e.g., DNA or RNA, encoding the TREM;

Maintaining the mammalian cell under conditions sufficient to express the TREM; and/or

Purifying the TREM from the mammalian host cell, e.g., according to the methods described herein.

14. The method of claim 12 or 13, wherein the mammalian host cell is selected from the group consisting of: non-human cells or cell lines, or human cells or cell lines, such as HEK293T cells (e.g., Freestyle 293-F cells), HT-1080 cells, PER. C6 cells, HKB-11 cells, CAP cells, HuH-7 cells, BHK 21 cells, MRC-S cells, MDCK cells, VERO cells, WI-38 cells, Chinese Hamster Ovary (CHO) cells, or MCF7 cells.

15. The method of any one of claims 12 to 14, wherein the purification step comprises one, two or all of the following steps, e.g. in the order:

(i) separating the nucleic acids from the cell debris to provide an RNA preparation;

(iii) TREM is separated from other RNA species in the small RNA preparation by affinity-based separation, e.g., sequence affinity-based separation.

16. The method of any one of claims 12-15, wherein the TREM comprises:

(i) an RNA sequence at least 80% identical to an RNA sequence encoded by a DNA sequence listed in table 1, or a fragment or functional fragment thereof; or

(ii) RNA sequences comprising the consensus sequences provided herein.

17. A method of making a tRNA effector molecule (TREM) composition, the method comprising:

(a) providing a mammalian host cell comprising an exogenous nucleic acid, e.g., DNA or RNA, encoding a TREM under conditions sufficient to express the TREM, and

(b) purifying the expressed TREM from the mammalian host cell to produce a TREM composition,

thereby preparing the TREM composition.

18. A method of making a pharmaceutical TREM composition, the method comprising:

combination of

a) A TREM, e.g., a purified TREM composition, e.g., a TREM composition prepared by a method described herein; and

b) pharmaceutically acceptable components, e.g., excipients,

thereby preparing a pharmaceutical TREM composition.

19. A method of making a purified tRNA effector molecule (TREM) pharmaceutical composition, the method comprising:

purifying the TREM from the mammalian host cell;

Formulating the purified TREM into a pharmaceutical composition, e.g. by combining said TREM with a pharmaceutical excipient,

thereby preparing the TREM pharmaceutical composition.

20. A method of making a TREM composition, the method comprising:

contacting a reaction mixture containing TREM with a reagent, such as a capture reagent or a separation reagent, comprising a nucleic acid sequence complementary to TREM;

thereby preparing a TREM composition.

21. A method of making a pharmaceutical composition, the method comprising:

a) providing a purified TREM composition, e.g., a purified TREM composition prepared by: culturing a mammalian host cell comprising DNA or RNA encoding said TREM under conditions sufficient to express the TREM, and purifying the expressed TREM from the host cell culture to produce a purified TREM composition,

b) providing a value, e.g., by assessing or testing one or more of the following characteristics of the purified TREM composition:

(i) purity of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%;

(iv) DNA, e.g., host cell DNA, of less than 1ng/ml, 5ng/ml, 10ng/ml, 15ng/ml, 20ng/ml, 25ng/ml, 30ng/ml, 35ng/ml, 40ng/ml, 50ng/ml, 60ng/ml, 70ng/ml, 80ng/ml, 90ng/ml or 100 ng/ml;

(v) less than 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% TREM fragments relative to full-length TREM;

(vi) low or absent endotoxin levels, e.g., a negative result as measured by a Limulus Amoebocyte Lysate (LAL) test;

(vii) in vitro translation activity, e.g., as measured by the assay described in example 15;

(x) Viral contamination, e.g., the composition or formulation is not present or is not detectable at a level of viral contamination;

c) optionally, formulating the purified TREM composition into a pharmaceutical product (e.g., combining the TREM composition with a pharmaceutical excipient) if the purified TREM composition meets the reference criterion for the one or more characteristics,

thereby preparing a pharmaceutical composition.

22. A pharmaceutical tRNA effector molecule (TREM) composition, comprising

(i) An RNA sequence at least 80% identical to an RNA sequence encoded by a DNA sequence listed in table 1, or a fragment or functional fragment thereof; or

(ii) RNA sequences comprising the consensus sequences provided herein.

23. A recombinant TREM composition that is at least 0.5g, 1g, 2g, 3g, 4g, 5g, 6g, 7g, 8g, 9g, 10g, 15g, 20g, 30g, 40g, 50g, 100g, 200g, 300g, 400g, or 500 g.

24. A recombinant TREM composition that is 0.5g to 500g, 0.5g to 400g, 0.5g to 300g, 0.5g to 200g, 0.5g to 100g, 0.5g to 50g, 0.5g to 40g, 0.5g to 30g, 0.5g to 20g, 0.5g to 10g, 0.5g to 9g, 0.5g to 8g, 0.5g to 7g, 0.5g to 6g, 0.5g to 5g, 0.5g to 4g, 0.5g to 3g, 0.5g to 2g, 0.5g to 1g, 1g to 500g, 2g to 500g, 5g to 500g, 10g to 500g, 20g to 500g, 30g to 500g, 40g to 500g, 50g to 500g, 100g to 500g, 300g to 500g, or 500 g.

25. A TREM composition comprising formula I_ZZZ(ii) a consensus sequence of (a),

R₀-R₁-R₂-R₃-R₄-R₅-R₆-R₇-R₈-R₉-R₁₀-R₁₁-R₁₂-R₁₃-R₁₄-R₁₅-R₁₆-R₁₇-R₁₈-R₁₉-R₂₀-R₂₁-R₂₂-R₂₃-R₂₄-R₂₅-R₂₆-R₂₇-R₂₈-R₂₉-R₃₀-R₃₁-R₃₂-R₃₃-R₃₄-R₃₅-R₃₆-R₃₇-R₃₈-R₃₉-R₄₀-R₄₁-R₄₂-R₄₃-R₄₄-R₄₅-R₄₆-[R₄₇]_x-R₄₈-R₄₉-R₅₀-R₅₁-R₅₂-R₅₃-R₅₄-R₅₅-R₅₆-R₅₇-R₅₈-R₅₉-R₆₀-R₆₁-R₆₂-R₆₃-R₆₄-R₆₅-R₆₆-R₆₇-R₆₈-R₆₉-R₇₀-R₇₁-R₇₂

wherein:

r is a ribonucleotide residue;

(i)_ZZZrepresents any one of twenty amino acids;

(ii) formula I corresponds to all species; and

(iii) x-1-271 (e.g., x-1-250, x-1-225, x-1-200, x-1-175, x-1-150, x-1-125, x-1-100, x-1-75, x-1-50, x-1-40, x-1-30, x-1-29, x-1-28, x-1-27, x-1-26, x-1-25, x-1-24, x-1-23, x-1-22, x-1-21, x-1-20, x-1-19, x-1-18, x-1-17, x-1-16, x-1-15, x-1-14, x-1-10, x-1-13, x-1-10, x-1-13, x-1-17, x-1-16, x-1-15, x-1-14, x-1-10, x-1-13, x-1-10, x-1-25, x-1-15, x-1, x-1-15, x-1, x-1, x-1, x-1, x-1, x-1, x-1, x, 1-1, x-1, x, 1, x-10-271, x-20-271, x-30-271, x-40-271, x-50-271, x-60-271, x-70-271, x-80-271, x-100-271, x-125-271, x-150-271, x-175-271, x-200-271, x-225-271, x-1, x-2, x-3, x-4, x-5, x-6, x-7, x-8, x-9, x-10, x-11, x-12, x-13, x-14, x-15, x-16, x-18, x-28, x-20, x-15, x-15, x-15, x-15, x-15, x-15, x-15, x-15, x, x-110, x-125, x-150, x-175, x-200, x-225, x-250, or x-271).

26. A TREM composition comprising formula II_ZZZ(ii) a consensus sequence of (a),

wherein:

r is a ribonucleotide residue;

(i)_ZZZrepresents any one of twenty amino acids;

(ii) formula II corresponds to a mammal; and

27. A TREM composition comprising formula III_ZZZ(ii) a consensus sequence of (a),

wherein:

r is a ribonucleotide residue;

(i)_ZZZrepresents any one of twenty amino acids;

(ii) formula III corresponds to human; and

28. A method of contacting a cell, tissue or subject with a TREM, the method comprising

Contacting the cell, tissue or subject with a purified TREM composition,

thereby contacting the cell, tissue or subject with the TREM.

29. A method of presenting TREM to a cell, tissue or subject having TREM, the method comprising

Contacting the cell, tissue or subject with a purified TREM composition,

thereby presenting the TREM to a cell, tissue, or subject.

30. A method of forming a cell, tissue or subject in contact with a TREM, the method comprising

Contacting the cell, tissue or subject with a purified TREM composition,

thereby forming a cell, tissue, or subject in contact with TREM.

31. A method of using TREM, the method comprising

Contacting the cell, tissue or subject with a purified TREM composition,

thereby using the TREM.

32. A method of applying TREM to a cell, tissue or subject, the method comprising

Contacting the cell, tissue or subject with a purified TREM composition,

thereby applying TREM to the cell, tissue or subject.

33. A method of exposing a cell, tissue or subject to TREM, the method comprising

Contacting the cell, tissue or subject with a purified TREM composition,

thereby exposing the cell, tissue or subject to TREM.

34. A method of forming a mixture of TREM and a cell, tissue or subject, the method comprising

Contacting the cell, tissue or subject with a TREM composition,

thereby forming a mixture of TREM and cells, tissue or subject.

35. A method of delivering TREM to a cell, tissue or subject, the method comprising:

providing a cell, tissue, or subject, and contacting the cell, tissue, or subject with a TREM composition, e.g., a purified TREM composition, e.g., a pharmaceutical TREM composition.

36. A method, such as an ex vivo method, of modulating metabolism, such as the translational capacity of an organelle, comprising:

providing a preparation of an organelle, e.g., a mitochondrion or chloroplast, and contacting the organelle with a pharmaceutical TREM composition.

37. A method of treating a subject, e.g., modulating metabolism, e.g., translational capacity of a cell, in a subject, the method comprising:

providing, e.g., administering to the subject an exogenous nucleic acid, e.g., DNA or RNA, encoding a TREM,

Thereby treating the subject.

38. A cell comprising a TREM prepared according to a method of preparing a TREM disclosed herein.

39. A cell comprising a TREM as disclosed herein.

40. A cell comprising an exogenous nucleic acid, the exogenous nucleic acid comprising:

a nucleic acid sequence, e.g., DNA or RNA, encoding a TREM, wherein the nucleic acid sequence comprises:

(i) a control region sequence;

(ii) a sequence encoding a modified TREM;

(iii) a sequence encoding more than one TREM;

(iv)tRNA^Meta sequence other than a sequence; or

(v) A promoter sequence comprising a Pol III recognition site, such as the U6 promoter, the 7SK promoter, or the H1 promoter, or a fragment thereof.

41. A reaction mixture comprising TREM and a reagent, such as a capture reagent or a separation reagent.

42. A bioreactor comprising a plurality of mammalian host cells described herein, the mammalian host cells comprising exogenous DNA or RNA.

43. A master cell bank comprising host cells, e.g., as described herein.

44. A method of assessing a composition of TREM, e.g., GMP-grade TREM (i.e., a TREM that conforms to cGMP and/or is prepared according to similar requirements), the method comprising obtaining a value for one or more of the following characteristics of the purified TREM composition:

(i) Purity of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%;

(iv) DNA, e.g., host cell DNA, of less than 1ng/ml, 5ng/ml, 10ng/ml, 15ng/ml, 20ng/ml, 25ng/ml, 30ng/ml, 35ng/ml, 40ng/ml, 50ng/ml, 60ng/ml, 70ng/ml, 80ng/ml, 90ng/ml or 100 ng/ml;

(v) less than 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% TREM fragments relative to full-length TREM;

(vi) low or absent endotoxin levels, e.g., a negative result as measured by a Limulus Amoebocyte Lysate (LAL) test;

(vii) in vitro translation activity, e.g., as measured by the assay described in example 15;

(ix) aseptically, e.g., the composition or formulation supports the growth of less than 100 viable microorganisms when tested under aseptic conditions, the composition or formulation meets the criteria of USP <71>, and/or the composition or formulation meets the criteria of USP <85> as described in cGMP guidelines for aseptically processed manufactured sterile pharmaceutical products; or

(x) Viral contamination, e.g., the composition or formulation is not present or no level of viral contamination is detected.

Background

trnas are complex RNA molecules that serve multiple functions, including initiation and elongation of proteins.

Disclosure of Invention

In one aspect, the disclosure provides a method of making a purified tRNA effector molecule (TREM) pharmaceutical composition, the method comprising:

providing a mammalian host cell comprising an exogenous nucleic acid, e.g., DNA or RNA, encoding the TREM;

maintaining the mammalian cell under conditions sufficient to express the TREM;

purifying the TREM from the mammalian host cell, e.g., according to the methods described herein; and is

Formulating the purified TREM into a pharmaceutical composition, e.g. by combining said TREM with a pharmaceutical excipient,

thereby preparing the TREM pharmaceutical composition.

In embodiments, the nucleic acid comprises RNA, which upon reverse transcription produces DNA that can be transcribed into TREM.

In embodiments, the nucleic acid comprises an RNA sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%) identical to an RNA sequence encoded by a DNA sequence listed in table 1, or a fragment or functional fragment thereof.

In embodiments, the nucleic acid comprises an RNA sequence comprising a consensus sequence, e.g., as provided herein, e.g., formula I_ZZZFormula II_ZZZOr formula III_ZZZWherein_ZZZRepresents any one of twenty amino acids: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine or valine.

In embodiments, the mammalian host cell is selected from the group consisting of: non-human cells or cell lines, or human cells or cell lines, such as HEK293T cells (e.g., Freestyle 293-F cells), HT-1080 cells, PER. C6 cells, HKB-11 cells, CAP cells, HuH-7 cells, BHK 21 cells, MRC-S cells, MDCK cells, VERO cells, WI-38 cells, Chinese Hamster Ovary (CHO) cells, or MCF7 cells.

In embodiments, the purification step comprises one, two or all of the following steps, e.g., in the order recited:

(i) separating the nucleic acids from the cell debris to provide an RNA preparation;

(iii) TREM is separated from other RNA species in the small RNA preparation by affinity-based separation, e.g., sequence affinity-based separation.

In one aspect, the invention features a method of making a tRNA effector molecule (TREM) composition, the method comprising:

(a) providing a host cell comprising an exogenous nucleic acid, e.g., DNA or RNA, encoding a TREM under conditions sufficient to express the TREM, and

(b) purifying the expressed TREM from the host cell culture to produce a TREM composition,

thereby preparing a TREM composition.

In embodiments, the TREM composition is a pharmaceutically acceptable composition.

In another aspect, the invention features a method of making a pharmaceutical TREM composition, the method including:

a) Providing a purified TREM composition, e.g., a purified TREM composition prepared by: culturing a mammalian host cell comprising DNA or RNA encoding said TREM under conditions sufficient to express the TREM, and purifying the expressed TREM from the host cell culture to produce a purified TREM composition,

b) providing a value, e.g., by assessing or testing a feature described herein (e.g., a feature related to identity (e.g., sequence), purity (e.g., a process impurity, such as a TREM fragment, a host cell protein, or a host cell DNA), activity (e.g., adaptor activity)),

thereby preparing the pharmaceutical TREM composition.

In another aspect, the invention features a method of making a pharmaceutical TREM composition, the method including:

combination of

a) A TREM, e.g., a purified TREM composition, e.g., a TREM composition prepared by a method described herein; and

b) pharmaceutically acceptable components, e.g., excipients,

Thereby preparing a pharmaceutical TREM composition.

In another aspect, the disclosure provides a composition comprising a purified tRNA effector molecule (TREM) (e.g., a purified TREM composition prepared according to a method described herein) comprising an RNA sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%) identical to an RNA sequence encoded by a DNA sequence listed in table 1, or a fragment or functional fragment thereof.

In one aspect, the disclosure provides compositions comprising a purified tRNA effector molecule (TREM) (e.g., a purified TREM composition prepared according to a method described herein), and methods of making the sameThe purified tRNA effector molecule comprises an RNA sequence that comprises a consensus sequence provided herein, e.g., formula I_ZZZFormula II_ZZZOr formula III_ZZZWherein_ZZZRepresents any of twenty amino acids: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine or valine.

In another aspect, the invention features a GMP-grade recombinant TREM composition (e.g., a TREM composition that conforms to cGMP and/or is prepared according to similar requirements) comprising an RNA sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%) identical to an RNA encoded by a DNA sequence listed in table 1, or a fragment or functional fragment thereof.

In another aspect, the invention features GMP-grade recombinant TREM compositions (e.g., a TREM composition that conforms to cGMP and/or is prepared according to similar requirements) that include an RNA sequence that includes a consensus sequence.

In one aspect, the invention features a TREM comprising a consensus sequence provided herein.

In one aspect, the invention features a composition that includes formula I_ZZZOf the consensus sequence of (1), wherein_ZZZRepresents any of the twenty amino acids and formula I corresponds to all species.

In one aspect, the invention features a composition that includes formula II_ZZZOf the consensus sequence of (1), wherein_ZZZRepresents any of the twenty amino acids and formula II corresponds to a mammal.

In one aspect, the invention features a composition that includes formula III_ZZZOf the consensus sequence of (1), wherein _ZZZRepresents any of the twenty amino acids and formula III corresponds to human.

In embodiments, ZZZ represents any one of the following amino acids: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine or valine.

In one aspect, the invention features a GMP-grade recombinant TREM composition comprising an RNA sequence comprising a consensus sequence provided herein.

In embodiments of any of the TREM compositions or pharmaceutical TREM compositions provided herein, the composition comprises one or more, e.g., a plurality of TREMs.

In an embodiment of any of the TREM compositions or pharmaceutical TREM compositions provided herein, the composition comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 TREMs.

In embodiments of any of the TREM compositions or pharmaceutical TREM compositions provided herein, the TREM composition (or an intermediate in the production of the TREM composition) comprises one or more of the following characteristics:

(i) purity of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%;

(ii) Host Cell Protein (HCP) contamination of less than 0.1ng/ml, 1ng/ml, 5ng/ml, 10ng/ml, 15ng/ml, 20ng/ml, 25ng/ml, 30ng/ml, 35ng/ml, 40ng/ml, 50ng/ml, 60ng/ml, 70ng/ml, 80ng/ml, 90ng/ml or 100 ng/ml;

(iv) DNA, e.g., host cell DNA, of less than 1ng/ml, 5ng/ml, 10ng/ml, 15ng/ml, 20ng/ml, 25ng/ml, 30ng/ml, 35ng/ml, 40ng/ml, 50ng/ml, 60ng/ml, 70ng/ml, 80ng/ml, 90ng/ml or 100 ng/ml;

(v) less than 0.1%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% fragments; (vi) low or absent endotoxin levels, e.g., a negative result as measured by a Limulus Amoebocyte Lysate (LAL) test;

(vii) in vitro translation activity, e.g., as measured by the assay described in example 15;

(ix) Aseptically, e.g., according to the cGMP guidelines for sterile pharmaceutical products, e.g., the composition or formulation supports the growth of less than 100 viable microorganisms when tested under sterile conditions, the composition or formulation meets the criteria of USP <71>, and/or the composition or formulation meets the criteria of USP <85 >; or

(x) Viral contamination, e.g., the composition or formulation is not present or no level of viral contamination is detected.

In another aspect, the invention features a cell that includes an exogenous nucleic acid that includes:

a nucleic acid sequence, e.g., DNA or RNA, encoding a TREM, wherein the nucleic acid sequence comprises:

(i) a control region sequence;

(ii) a sequence encoding a modified TREM;

(iii) a sequence encoding more than one TREM;

(iv)tRNA^Meta sequence other than a sequence; or

(v) A promoter sequence comprising a Pol III recognition site, such as the U6 promoter, the 7SK promoter, or the H1 promoter, or a fragment thereof.

In one aspect, the invention features a method of modulating a tRNA pool in a cell, the method comprising:

providing a purified TREM composition, and contacting the cell with the TREM composition,

thereby modulating the tRNA pool in the cell.

In another aspect, the invention features a method of delivering TREM to a cell, tissue, or subject, the method including:

providing a cell, tissue or subject, and contacting the cell, tissue or subject with a TREM composition comprising the TREM, e.g., a pharmaceutical TREM composition comprising the TREM.

In another aspect, the invention features a method of treating a subject, e.g., modulating metabolism, e.g., translational capacity of a cell, in a subject, the method including:

providing, e.g., administering to the subject an exogenous nucleic acid, e.g., DNA or RNA, encoding a TREM, thereby treating the subject.

In embodiments of any of the methods disclosed herein, the TREM composition is prepared from:

providing a mammalian host cell comprising an exogenous nucleic acid, e.g., DNA or RNA, encoding the TREM;

maintaining the mammalian cell under conditions sufficient to express the TREM; and/or

Purifying the TREM from the mammalian host cell, e.g., according to the methods described herein.

In embodiments of any of the methods disclosed herein, the mammalian host cell is a non-human cell or cell line, or a human cell or cell line selected from the group consisting of: HEK293T cells (e.g., Freestyle293-F cells), HT-1080 cells, PER. C6 cells, HKB-11 cells, CAP cells, HuH-7 cells, BHK 21 cells, MRC-S cells, MDCK cells, VERO cells, WI-38 cells, Chinese Hamster Ovary (CHO) cells, or MCF7 cells.

In embodiments of any of the methods disclosed herein, the purifying step comprises one, two, or all of the following steps, e.g., in the order recited:

(i) separating the nucleic acids from the cell debris to provide an RNA preparation;

(iii) TREM is separated from other RNA species by affinity-based separation, e.g., sequence affinity-based separation.

In embodiments of any of the methods disclosed herein, TREM comprises:

(i) an RNA sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%) identical to an RNA sequence encoded by a DNA sequence listed in table 1, or a fragment or functional fragment thereof; or

(ii) RNA sequences comprising the consensus sequences provided herein.

In one aspect, the disclosure provides a method of making a purified tRNA effector molecule (TREM) pharmaceutical composition, the method comprising:

providing an insect host cell comprising an exogenous nucleic acid, e.g., DNA or RNA, encoding a TREM;

Maintaining the insect host cell under conditions sufficient to express a TREM;

purifying TREM from the insect host cell, e.g., according to the methods described herein; and is

Formulating the purified TREM into a pharmaceutical composition, e.g. by combining said TREM with a pharmaceutical excipient,

thereby preparing the TREM pharmaceutical composition.

In embodiments, the insect host cell is selected from the group consisting of: insect cells or cell lines, for example, Sf9 cells or cell lines.

In embodiments, the purification step comprises one, two or all of the following steps, e.g., in the order recited:

(i) separating the nucleic acids from the proteins to provide an RNA preparation;

(iii) TREM is separated from other RNA species in the small RNA preparation by affinity-based separation, e.g., sequence affinity.

In one aspect, the disclosure provides a method of making a purified tRNA effector molecule (TREM) pharmaceutical composition, the method comprising:

providing a yeast host cell comprising an exogenous nucleic acid, e.g., DNA or RNA, encoding a TREM;

Maintaining the yeast host cell under conditions sufficient to express a TREM;

purifying TREM from the yeast host cell, e.g., according to the methods described herein; and is

Formulating the purified TREM into a pharmaceutical composition, e.g. by combining said TREM with a pharmaceutical excipient,

thereby preparing the TREM pharmaceutical composition.

In embodiments, the yeast host cell is selected from the group consisting of: yeast cells or cell lines, for example Saccharomyces cerevisiae or Schizosaccharomyces pombe cells or cell lines.

In embodiments, the purification step comprises one, two or all of the following steps, e.g., in the order recited:

(i) separating the nucleic acids from the proteins to provide an RNA preparation;

(iii) TREM is separated from other RNA species in the small RNA preparation by affinity-based separation, e.g., sequence affinity.

tRNA-based effector molecules (TREMs) as disclosed herein are complex molecules that can mediate a variety of cellular processes. The pharmaceutical TREM composition may be administered to a cell, tissue, or subject to modulate these functions, e.g., in vitro or in vivo. Disclosed herein are TREM compositions, formulations, methods of making TREM compositions and formulations, and methods of using TREM compositions and formulations.

Additional features of any of the foregoing TREM compositions, formulations, methods of making TREM compositions and formulations, and methods of using TREM compositions and formulations include one or more of the examples set forth below.

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalent embodiments are intended to be encompassed by the embodiments listed below.

Illustrative examples

1. A method of making a purified tRNA effector molecule (TREM) pharmaceutical composition, the method comprising:

providing a mammalian host cell comprising an exogenous nucleic acid, e.g., DNA or RNA, encoding the TREM;

maintaining the mammalian host cell under conditions sufficient to express a TREM;

purifying the TREM from the mammalian host cell, e.g., according to the methods described herein; and

formulating the purified TREM into a pharmaceutical composition, e.g. by combining said TREM with a pharmaceutical excipient,

thereby preparing the TREM pharmaceutical composition.

2. A method of making a tRNA effector molecule (TREM) composition, the method comprising:

(a) providing a mammalian host cell comprising an exogenous nucleic acid, e.g., DNA or RNA, encoding a TREM under conditions sufficient to express the TREM, and

(b) Purifying the expressed TREM from the mammalian host cell to produce a TREM composition,

thereby preparing the TREM composition.

3. The method of example 2, formulating said TREM composition into a pharmaceutical composition, e.g., by combining said TREM with a pharmaceutical excipient,

4. a method of making a pharmaceutical TREM composition, the method comprising:

combination of

a) A TREM, e.g., a purified TREM composition, e.g., a TREM composition prepared by a method described herein; and

b) pharmaceutically acceptable components, e.g., excipients,

thereby preparing a pharmaceutical TREM composition.

5. The method of claim 4, wherein the TREM is purified from a mammalian host cell, e.g., according to a method described herein.

6. A method of making a purified tRNA effector molecule (TREM) pharmaceutical composition, the method comprising:

purifying the TREM from the mammalian host cell;

formulating the purified TREM into a pharmaceutical composition, e.g. by combining said TREM with a pharmaceutical excipient,

thereby preparing the TREM pharmaceutical composition.

7. The method of claim 5 or 6, wherein the mammalian host cell comprises an exogenous nucleic acid, such as DNA or RNA, encoding a TREM.

8. The method of any one of embodiments 1-7, wherein the purification step comprises one, two, or all of the following steps, e.g., in the order recited:

(i) separating the nucleic acids from the proteins to provide an RNA preparation;

(iii) TREM is separated from other RNA species in the small RNA preparation by affinity-based separation, e.g., sequence affinity.

9. The method of embodiment 8, comprising step (i).

10. The method of embodiment 8 or 9, comprising step (ii).

11. The method of any one of embodiments 8 to 10, comprising step (iii).

12. The method of any of embodiments 8 or 10-11, comprising performing:

step (i) preceding step (ii).

13. The method of any one of embodiments 8 or 11-12, comprising performing step (ii) prior to step (iii).

14. The method of any one of embodiments 8-13, wherein (i) comprises extracting nucleic acids from proteins.

15. The method of any one of embodiments 8-14, wherein (i) comprises a phenol/chloroform extraction.

16. The method of any one of embodiments 8-10 or 12-15, wherein (ii) comprises separating RNA of less than a first size scale from RNA of a larger second size scale.

17. The method of embodiment 16, wherein the first size level is less than 200 nt.

18. The method of any one of embodiments 8 or 9-16, wherein (ii) comprises performing salt precipitation to enrich for less than 200nt of RNA.

19. The method of embodiment 18, wherein the salt comprises LiCl.

20. The method of any one of embodiments 8-10 or 12-19, wherein (ii) further comprises performing a desalting or buffer exchange step.

21. The method of any one of embodiments 8 or 11-20, wherein (iii) comprises performing affinity-based separation to enrich TREM.

22. The method of embodiment 21, wherein affinity-based separation comprises sequence-based separation, e.g., using a probe comprising a sequence that binds to TREM.

23. The method of any one of the preceding embodiments, wherein the TREM composition is a pharmaceutically acceptable composition.

24. The method of any one of embodiments 1-3 or 7-23, comprising introducing exogenous DNA or RNA into the mammalian host cell.

25. The method of any one of embodiments 1-3 or 7-24, wherein the nucleic acid comprises DNA that expresses TREM post-transcriptionally.

26. The method of any one of embodiments 1-3 or 7-25, wherein the nucleic acid comprises RNA that upon reverse transcription produces DNA that can be transcribed to provide a TREM.

27. The method of any one of the preceding embodiments, wherein the TREM recognizes a stop codon.

28. The method of claim 27, wherein the TREM mediates acceptance and incorporation of amino acids.

29. The method of any one of embodiments 1-27, wherein the TREM does not recognize a stop codon.

30. The method of any one of embodiments 1-29, wherein the TREM comprises:

(ii) RNA sequences comprising the consensus sequences provided herein.

31. The method of any one of the preceding embodiments, wherein the TREM composition comprises a TREM fragment, e.g., as described herein, optionally wherein the TREM fragment is produced in vivo in the host cell.

32. The method of embodiment 31, wherein the TREM fragment is produced by fragmenting an expressed TREM after the cell produces a TREM, e.g., a TREM produced by the host cell is fragmented after release or purification from the host cell, e.g., the TREM is fragmented ex vivo.

33. The method of any one of the preceding embodiments, wherein the method results in increased production of total endogenous tRNA and TREM in the host cell, e.g., by at least 2.2, 2.5, 3, 4, 5, 6, 7, 8, 9, 10, or 20-fold (e.g., as measured by the assay described in any of examples 7-11), e.g., as compared to a reference cell, e.g., a similar cell, but without engineering or modification to express TREM.

34. The method of embodiment 33, wherein the method results in a 2.2 to 20-fold, 2.2 to 15-fold, 2.2 to 10-fold, 2.2 to 9-fold, 2.2 to 8-fold, 2.2 to 7-fold, 2.2 to 6-fold, 2.2 to 5-fold, 2.2 to 4-fold, 2.2 to 3-fold, 2.2 to 2.5-fold, 2.5 to 20-fold, 3 to 20-fold, 4 to 20-fold, 5 to 20-fold, 6 to 20-fold, 7 to 20-fold, 8 to 20-fold, 9 to 20-fold, 10 to 20-fold, or 15 to 20-fold increase in TREM yield and/or tRNA yield.

35. The method of any one of the preceding embodiments, wherein the method results in a detectable level of TREM in the host cell, e.g., as measured by the assay of any one of examples 7-11.

36. The method of any one of the preceding embodiments, wherein the host cell is capable of post-transcriptional modification of the TREM.

37. The method of any one of the preceding embodiments, wherein the host cell is capable of post-transcriptional modification of the TREM, e.g., a post-transcriptional modification selected from table 2.

38. The method of any one of the preceding embodiments, wherein the host cell has been modified to modulate, e.g., increase its ability to provide a TREM post-transcriptional modification, e.g., a post-transcriptional modification selected from table 2, e.g., the host cell has been modified to provide, increase or decrease expression of a gene, e.g., a gene encoding an enzyme from table 2, or a gene encoding an enzyme having nuclease activity (e.g., endonuclease activity or ribonuclease activity), e.g., or one or more of Dicer, angiogenin, rnase A, RNA enzyme P, RNA enzyme Z, Rny1 or PrrC.

39. The method of any one of the preceding embodiments, wherein the host cell is a mammalian cell capable of post-transcriptional modification of the TREM, e.g., a post-transcriptional modification selected from table 2.

40. The method of any one of the preceding embodiments, wherein the host cell comprises a cell selected from the group consisting of: HEK293T cells (e.g., Freestyle 293-F cells), HT-1080 cells, PER. C6 cells, HKB-11 cells, CAP cells, HuH-7 cells, BHK 21 cells, MRC-S cells, MDCK cells, VERO cells, WI-38 cells, Chinese Hamster Ovary (CHO) cells, or MCF7 cells.

41. The method of any one of the preceding embodiments, wherein the host cell comprises a HeLa cell, a HEK293 cell, an HT-1080 cell, a PER. C6 cell, a HKB-11 cell, a CAP cell, or a HuH-7 cell.

42. The method of any one of the preceding embodiments, wherein the host cell has increased oncogene expression, e.g., Ras, c-myc, or c-jun.

43. The method of any one of the preceding embodiments, wherein the host cell has reduced tumor suppressor expression, e.g., reduced expression of p53 or Rb.

44. The method of any one of the preceding embodiments, wherein the host cell has increased expression of RNA polymerase iii (RNA Pol iii).

45. The method of any one of the preceding embodiments, wherein the host cell has increased tRNA ^MetExpression, e.g. increased tRNA^iMetOr tRNA^eMetAnd (4) expressing.

46. The method of any one of the preceding embodiments, comprising culturing the host cell in a medium that promotes cell hyperproliferation (e.g., that promotes signaling pathways that are amplified in cancer cells).

47. The method of any one of the preceding embodiments, comprising culturing the host cell in a growth-promoting medium, e.g., a medium comprising or supplemented with: one or a combination of growth factors, cytokines, or hormones, for example, one or a combination of serum (e.g., Fetal Bovine Serum (FBS)), Fibroblast Growth Factor (FGF), Epidermal Growth Factor (EGF), insulin-like growth factor (IGF), transforming growth factor beta (TGFb), platelet-derived growth factor (PDGF), Hepatocyte Growth Factor (HGF), or Tumor Necrosis Factor (TNF).

48. The method of any one of the preceding embodiments, comprising culturing the host cell in a medium that facilitates post-transcriptional processing of, for example, TREM.

49. The method of any one of the preceding embodiments, comprising culturing the host cell under certain conditions, such as a medium that promotes overexpression or overactivation of enzymes involved in post-transcriptional processing, for example, under conditions that promote:

a) Removal of the 5' leader sequence, for example by rnase P;

b) 3' tail exonuclease activity, e.g., rnase II, PNP, rnase PH, or rnase T activity;

c) adding CCA at the 3' end, e.g., by nucleotidyl transferase;

d) intron splicing, for example by one or more (e.g., all) of: a splicing endonuclease, cyclic phosphodiesterase, adenylyltransferase, ligase, or 2' phosphotransferase;

e) modifying, e.g., by modifying an enzyme, e.g., an enzyme having one or more of the following enzymatic activities:

(i) adenosine A₃₄To inosine I₃₄Deamination;

(ii) adenosine m¹A₅₈Methylation of (a);

(iii) proceed to ncm⁵Um₃₄ or ncm⁵s²U₃₄Modifying;

(iv) carry out ct⁶A modification; isoamylation of i⁶A₃₇Modifying; a. the₃₇To i⁶A₃₇Modifying; or

(v) The modifications listed in table 2 were performed; or

f) A synthetase involved in amino acid loading.

50. The method of any one of the preceding embodiments, comprising culturing the host cell in a medium with excess nutrients, e.g., not nutrient limited.

51. The method of any one of the preceding embodiments, comprising culturing the host cell in a medium that promotes expression of Mck1 and/or Kns1, e.g., increases Mck1 and/or Kns1 expression and/or activity.

52. The method of any one of the preceding embodiments, wherein the host cell has increased expression and/or activity of Trm 1.

53. The method of any one of the preceding embodiments, wherein the host cell has reduced activity of Maf1, e.g., by phosphorylation of serine at position 45 of Maf1, e.g., Maf 1.

54. The method of example 53, wherein a decrease in Maf1 activity results in an increase in TREM production.

55. The method of example 53 or 54, wherein the activity of Maf1 can be reduced by: introduction of a pseudo-phosphorylated Maf1 mutant e.g., a mutant having a serine to aspartate mutation at position 45 (S45D); or over-activating CK2/TORC1, for example, which phosphorylates Maf 1.

56. The method of any one of the preceding embodiments, further comprising measuring one or more of the following characteristics of the TREM composition (or an intermediate in the production of a TREM composition):

(i) purity of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%;

(iv) DNA, e.g., host cell DNA, of less than 1ng/ml, 5ng/ml, 10ng/ml, 15ng/ml, 20ng/ml, 25ng/ml, 30ng/ml, 35ng/ml, 40ng/ml, 50ng/ml, 60ng/ml, 70ng/ml, 80ng/ml, 90ng/ml or 100 ng/ml;

(v) less than 0.1%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% fragments; (vi) low or absent endotoxin levels, e.g., a negative result as measured by a Limulus Amoebocyte Lysate (LAL) test;

(vii) in vitro translation activity, e.g., as measured by the assay described in example 15;

(x) Viral contamination, e.g., the composition or formulation is not present or no level of viral contamination is detected.

57. The method of embodiment 56, further comprising comparing the measured value to a reference value or standard.

58. The method of embodiment 57, further comprising, in response to the comparing, adjusting the TREM composition to:

(i) increasing the purity of the TREM composition;

(ii) reducing the amount of HCP in the composition;

(iii) reducing the amount of DNA in the composition;

(iv) reducing the amount of fragments in the composition;

(v) reducing the amount of endotoxin in the composition;

(vi) increasing the in vitro translation activity of the composition;

(vii) increasing the TREM concentration of the composition; or

(viii) Increasing the sterility of the composition.

59. A method of making a TREM composition, the method comprising:

contacting a reaction mixture containing TREM with a reagent, such as a capture reagent or a separation reagent, comprising a nucleic acid sequence complementary to TREM;

thereby preparing a TREM composition.

60. The method of embodiment 59, further comprising denaturing the TREM, e.g., prior to hybridization to the capture reagent.

61. The method of example 59, further comprising renaturing the TREM, e.g., after hybridization and/or release from the capture reagent.

62. The method of any one of embodiments 59-61, further wherein a single capture reagent is used, e.g., to prepare a TREM composition in which at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of the TREMs have a sequence complementary to the capture reagent.

63. The method of any one of embodiments 59-61, further wherein a plurality of capture reagents are used, e.g., to prepare a TREM composition having a plurality of different TREMs.

64. A method of making a pharmaceutical composition, the method comprising:

b) providing a value, e.g., by assessing or testing one or more of the following characteristics of the purified TREM composition:

(i) purity of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%;

(iv) DNA, e.g., host cell DNA, of less than 1ng/ml, 5ng/ml, 10ng/ml, 15ng/ml, 20ng/ml, 25ng/ml, 30ng/ml, 35ng/ml, 40ng/ml, 50ng/ml, 60ng/ml, 70ng/ml, 80ng/ml, 90ng/ml or 100 ng/ml;

(v) less than 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% TREM fragments relative to full-length TREM;

(vi) low or absent endotoxin levels, e.g., a negative result as measured by a Limulus Amoebocyte Lysate (LAL) test;

(vii) in vitro translation activity, e.g., as measured by the assay described in example 15;

(x) Viral contamination, e.g., the composition or formulation is not present or no level of viral contamination is detected.

c) Optionally, formulating the purified TREM composition into a pharmaceutical product (e.g., combining the TREM composition with a pharmaceutical excipient) if the purified TREM composition meets the reference criterion for the one or more characteristics,

thereby preparing a pharmaceutical composition.

65. The method of embodiment 64, further comprising comparing the measured value to a reference value or standard.

66. The method of embodiment 65, further comprising, in response to the comparing, adjusting the composition to:

(i) increasing the purity of the TREM composition;

(ii) reducing the amount of HCP in the composition;

(iii) reducing the amount of DNA in the composition;

(iv) reducing the amount of fragments in the composition;

(v) reducing the amount of endotoxin in the composition;

(vi) Increasing the in vitro translation activity of the composition;

(vii) increasing the TREM concentration of the composition; or

(viii) Increasing the sterility of the composition.

67. A composition comprising a purified tRNA effector molecule (TREM) (e.g., a purified TREM composition prepared according to a method described herein), the purified tRNA effector molecule comprising:

(ii) RNA comprising the consensus sequences provided herein, and

optionally, the RNA sequence has less than 100% identity to an RNA sequence encoded by a DNA sequence listed in table 1.

68. A GMP-grade recombinant TREM composition (e.g., a TREM composition that conforms to cGMP and/or is prepared according to similar requirements), comprising:

(ii) RNA comprising the consensus sequences provided herein, and

optionally, the RNA sequence has less than 100% identity to an RNA sequence encoded by a DNA sequence listed in table 1.

69. A pharmaceutical tRNA effector molecule (TREM) composition comprising

(i) An RNA sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%) identical to an RNA sequence encoded by a DNA sequence listed in table 1, or a fragment or functional fragment thereof; or

(ii) RNA comprising the consensus sequences provided herein, and

optionally, the RNA sequence has less than 100% identity to an RNA sequence encoded by a DNA sequence listed in table 1.

70. The pharmaceutical TREM composition of claim 69, comprising a purified tRNA effector molecule (TREM) (e.g., a purified TREM composition prepared according to a method described herein).

71. The composition or pharmaceutical composition of any one of embodiments 67-70, wherein the TREM is prepared as described in any one of embodiments 1-66.

72. The composition or pharmaceutical composition of any one of embodiments 67-70, wherein said TREM comprises one or more post-transcriptional modifications listed in Table 2.

73. The composition or pharmaceutical composition of embodiment 72, wherein the TREM comprises one or more post-transcriptional modifications listed in Table 2.

74. A recombinant TREM composition of at least 0.5g, 1g, 2g, 3g, 4g, 5g, 6g, 7g, 8g, 9g, 10g, 15g, 20g, 30g, 40g, 50g, 100g, 200g, 300g, 400g, or 500 g.

75. A recombinant TREM composition that is 0.5g to 500g, 0.5g to 400g, 0.5g to 300g, 0.5g to 200g, 0.5g to 100g, 0.5g to 50g, 0.5g to 40g, 0.5g to 30g, 0.5g to 20g, 0.5g to 10g, 0.5g to 9g, 0.5g to 8g, 0.5g to 7g, 0.5g to 6g, 0.5g to 5g, 0.5g to 4g, 0.5g to 3g, 0.5g to 2g, 0.5g to 1g, 1g to 500g, 2g to 500g, 5g to 500g, 10g to 500g, 20g to 500g, 30g to 500g, 40g to 500g, 50g to 500g, 100g to 500g, 200g to 500g, 300g to 500g, or 500 g.

76. A TREM composition comprising formula I_ZZZ(ii) a consensus sequence of (a),

R₀-R_1-R₂-R₃-R₄-R₅-R₆-R₇-R₈-R₉-R₁₀-R₁₁-R₁₂-R₁₃-R₁₄-R₁₅-R₁₆-R₁₇-R₁₈-R₁₉-R₂₀-R₂₁-R₂₂-R₂₃-R₂₄-R₂₅-R₂₆-R₂₇-R₂₈-R₂₉-R₃₀-R₃₁-R₃₂-R₃₃-R₃₄-R₃₅-R₃₆-R₃₇-R₃₈-R₃₉-R₄₀-R₄₁-R₄₂-R₄₃-R₄₄-R₄₅-R₄₆-[R₄₇]_x-R₄₈-R₄₉-R₅₀-R₅₁-R₅₂-R₅₃-R₅₄-R₅₅-R₅₆-R₅₇-R₅₈-R₅₉-R₆₀-R₆₁-R₆₂-R₆₃-R₆₄-R₆₅-R₆₆-R₆₇-R₆₈-R₆₉-R₇₀-R₇₁-R₇₂

wherein:

r is a ribonucleotide residue;

(i)_ZZZrepresents any one of twenty amino acids;

(ii) formula I corresponds to all species; and

77. A TREM composition comprising formula II_ZZZ(ii) a consensus sequence of (a),

wherein:

r is a ribonucleotide residue;

(i)_ZZZrepresents any one of twenty amino acids;

(ii) formula II corresponds to a mammal; and

78. A TREM composition comprising formula III_ZZZ(ii) a consensus sequence of (a),

wherein:

r is a ribonucleotide residue;

(i)_ZZZrepresents any one of twenty amino acids;

(ii) formula III corresponds to human; and

79. The composition or pharmaceutical composition of any one of embodiments 67-78, wherein the composition comprises one or more, e.g., a plurality of TREMs.

80. The composition or pharmaceutical composition of any one of embodiments 67-79, wherein the composition comprises one or more unique TREM, e.g., one or more TREM comprising different anti-codon sequences.

81. The composition or pharmaceutical composition of any one of embodiments 67-80, wherein the composition comprises one or more unique TREMs, e.g., a TREM that recognizes a different codon.

82. The composition or pharmaceutical composition of any one of embodiments 67-81, wherein the TREM composition (or an intermediate in the production of a TREM composition) comprises one or more of the following characteristics:

(i) purity of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%;

(iv) DNA, e.g., host cell DNA, of less than 1ng/ml, 5ng/ml, 10ng/ml, 15ng/ml, 20ng/ml, 25ng/ml, 30ng/ml, 35ng/ml, 40ng/ml, 50ng/ml, 60ng/ml, 70ng/ml, 80ng/ml, 90ng/ml or 100 ng/ml;

(v) less than 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% TREM fragments relative to full-length TREM;

(vi) low or absent endotoxin levels, e.g., a negative result as measured by a Limulus Amoebocyte Lysate (LAL) test;

(vii) in vitro translation activity, e.g., as measured by the assay described in example 15;

(x) Viral contamination, e.g., the composition or formulation is not present or no level of viral contamination is detected.

83. A method of modulating a tRNA pool in a cell, the method comprising: providing a purified TREM composition, and contacting the cell with the TREM composition, thereby modulating the tRNA pool in the cell.

84. A method of contacting a cell, tissue, or subject with a TREM, the method comprising contacting the cell, tissue, or subject with a purified TREM composition, thereby contacting a cell, tissue, or subject with the TREM.

85. A method of presenting TREM to a cell, tissue or subject having TREM, the method comprising

Contacting the cell, tissue or subject with the purified TREM composition, thereby presenting the TREM to the cell, tissue or subject.

86. A method of forming a cell, tissue or subject in contact with a TREM, the method comprising

Contacting the cell, tissue or subject with the purified TREM composition, thereby forming a cell, tissue or subject contacted with TREM.

87. A method of using TREM, comprising contacting the cell, tissue or subject with a purified TREM composition, thereby using the TREM.

88. A method of applying TREM to a cell, tissue or subject, the method comprising contacting the cell, tissue or subject with a purified TREM composition, thereby applying TREM to the cell, tissue or subject.

89. A method of exposing a cell, tissue or subject to TREM, the method comprising contacting the cell, tissue or subject with a purified TREM composition, thereby exposing the cell, tissue or subject to TREM.

90. A method of forming a mixture of TREM and a cell, tissue or subject, the method comprising

Contacting the cell, tissue or subject with a TREM composition, thereby forming a mixture of TREM and cell, tissue or subject.

91. A method of delivering TREM to a cell, tissue or subject, the method comprising:

providing a cell, tissue, or subject, and contacting the cell, tissue, or subject with a TREM composition, e.g., a purified TREM composition, e.g., a pharmaceutical TREM composition.

92. A method, such as an ex vivo method, of modulating metabolism, such as the translational capacity of an organelle, comprising:

providing a preparation of an organelle, e.g., a mitochondrion or chloroplast, and contacting the organelle with a pharmaceutical TREM composition.

93. A method of treating a subject, e.g., modulating metabolism, e.g., translational capacity of a cell, in a subject, the method comprising:

providing, e.g., administering to the subject an exogenous nucleic acid, e.g., DNA or RNA, encoding a TREM,

thereby treating the subject.

94. The method of any one of embodiments 83-93, wherein the TREM composition is prepared as described in any one of embodiments 1-66 or the TREM comprises the composition provided in any one of embodiments 67-82.

95. The method of any one of embodiments 83-93, wherein the TREM composition is prepared by:

providing a mammalian host cell comprising an exogenous nucleic acid, e.g., DNA or RNA, encoding the TREM;

maintaining the mammalian cell under conditions sufficient to express the TREM; and/or

Purifying the TREM from the mammalian host cell, e.g., according to the methods described herein.

96. The method of embodiment 95, wherein the mammalian host cell is selected from the group consisting of: non-human cells or cell lines, or human cells or cell lines, such as HEK293T cells (e.g., Freestyle 293-F cells), HT-1080 cells, PER. C6 cells, HKB-11 cells, CAP cells, HuH-7 cells, BHK 21 cells, MRC-S cells, MDCK cells, VERO cells, WI-38 cells, Chinese Hamster Ovary (CHO) cells, or MCF7 cells.

97. The method of any one of embodiments 95-96, wherein the purification step comprises one, two, or all of the following steps, e.g., in the order recited:

(i) separating the nucleic acids from the cell debris to provide an RNA preparation;

(iii) TREM is separated from other RNA species in the small RNA preparation by affinity-based separation, e.g., sequence affinity-based separation.

98. The method of any one of embodiments 83-97, wherein the TREM comprises:

(ii) RNA comprising the consensus sequences provided herein, and

optionally, the RNA sequence has less than 100% identity to an RNA sequence encoded by a DNA sequence listed in table 1.

99. The method of any one of embodiments 83-98, wherein the method is an in vitro method, e.g., a cell or tissue is contacted with the TREM composition in vitro.

100. The method of any one of embodiments 83-98, wherein the method is an ex vivo method, e.g., a cell or tissue is contacted with the TREM composition ex vivo, and optionally, the contacted cell or tissue is introduced, e.g., administered, into a subject, e.g., a subject from which the cell or tissue is derived, or a different subject.

101. The method of any one of embodiments 83-98, wherein the method is an in vivo method, e.g., a subject or a tissue or cell of a subject is contacted with the TREM composition in vivo.

102. The method of any one of embodiments 99-101, comprising contacting the TREM composition, e.g., a pharmaceutical TREM composition, with a cell.

103. The method of any one of embodiments 99-101, comprising contacting the TREM composition, e.g., a pharmaceutical TREM composition, with a tissue.

104. The method of any one of embodiments 99-100 or 102, comprising administering a TREM composition, e.g., a pharmaceutical TREM composition, to a subject.

105. The method of any of embodiments 100 or 104, wherein the TREM composition is administered with a carrier or delivery agent, such as a liposome, a polymer (e.g., a polymer conjugate), a particle, a microsphere, a microparticle, or a nanoparticle.

106. The method of any one of embodiments 99-105, wherein the cell is cancerous.

107. The method of any one of embodiments 99-105, wherein the cell is non-cancerous.

108. The method of any one of embodiments 99-102 or 104-107, wherein the cell or tissue comprises:

muscle cells or tissues (e.g., skeletal muscle cells or tissues, smooth muscle cells or tissues, or cardiac muscle cells or tissues),

epithelial cells or tissues;

connective cells or tissues (e.g., adipocytes or tissues, osteocytes or tissues, or blood cells), or

A neural cell or tissue (e.g., a sensory neuron, a motor neuron, or an interneuron).

109. The method of any one of embodiments 99-108, wherein the method comprises administering to the subject the cells contacted with the TREM composition ex vivo or in vitro.

110. A cell comprising a TREM prepared as described in any one of examples 1-66.

111. A cell comprising a TREM as described in any one of embodiments 67-82.

112. A cell comprising an exogenous nucleic acid, the exogenous nucleic acid comprising:

a nucleic acid sequence, e.g., DNA or RNA, encoding a TREM, wherein the nucleic acid sequence comprises:

(i) A control region sequence;

(ii) a sequence encoding a modified TREM;

(iii) a sequence encoding more than one TREM;

(iv)tRNA^Meta sequence other than a sequence; or

(v) A promoter sequence comprising a Pol III recognition site, such as the U6 promoter, the 7SK promoter, or the H1 promoter, or a fragment thereof.

113. The method of any one of embodiments 111-112, wherein the host cell is capable of post-transcriptional modification of the TREM.

114. The method of any one of embodiments 111-113, wherein the host cell is capable of post-transcriptional modification of the TREM, e.g., a post-transcriptional modification selected from table 2.

115. The method of any one of embodiments 111-114, wherein the host cell has been modified to modulate, e.g., increase its ability to provide a TREM post-transcriptional modification, e.g., a post-transcriptional modification selected from table 2, e.g., the host cell has been modified to provide, increase or decrease expression of a gene, e.g., a gene encoding an enzyme from table 2, or a gene encoding an enzyme having nuclease activity (e.g., endonuclease activity or ribonuclease activity), e.g., or one or more of dicer, angiogenin, rnase A, RNA, enzyme P, RNA, enzyme Z, Rny1, or PrrC.

116. The method of any one of embodiments 111-115, wherein the host cell is a mammalian cell capable of post-transcriptional modification of the TREM, e.g., selected from table 2.

117. The method of any one of embodiments 111-116, wherein the host cell comprises a cell or cell line selected from the group consisting of: HEK293T cells (e.g., Freestyle 293-F cells), HT-1080 cells, PER. C6 cells, HKB-11 cells, CAP cells, HuH-7 cells, BHK 21 cells, MRC-S cells, MDCK cells, VERO cells, WI-38 cells, Chinese Hamster Ovary (CHO) cells, or MCF7 cells.

118. The method of any one of embodiments 111-117, wherein the host cell comprises a cell or cell line selected from the group consisting of: HeLa cells, HEK293 cells, HT-1080 cells, PER. C6 cells, HKB-11 cells, CAP cells, or HuH-7 cells.

119. The method of any one of embodiments 111-1118, wherein the host cell has increased expression of an oncogene, such as Ras, c-myc, or c-jun.

120. The method of any one of embodiments 111-119, wherein the host cell has reduced expression of a tumor suppressor, e.g., reduced expression of p53 or Rb.

121. The method of any one of embodiments 111-120, wherein the host cell has increased expression of RNA polymerase III (RNA Pol III).

122. The method of any one of embodiments 111-121, wherein the host cell has increased tRNA^MetExpression, e.g. increased tRNA^iMetOr tRNA^eMetAnd (4) expressing.

123. The method of any one of embodiments 111-122, comprising culturing the host cell in a medium that promotes cell hyperproliferation (e.g., that promotes a signaling pathway that is amplified in cancer cells).

124. The method of any one of embodiments 111-123, comprising culturing the host cell in a growth-promoting medium, e.g., a medium comprising or supplemented with: one or a combination of growth factors, cytokines, or hormones, for example, one or a combination of serum (e.g., Fetal Bovine Serum (FBS)), Fibroblast Growth Factor (FGF), Epidermal Growth Factor (EGF), insulin-like growth factor (IGF), transforming growth factor beta (TGFb), platelet-derived growth factor (PDGF), Hepatocyte Growth Factor (HGF), or Tumor Necrosis Factor (TNF).

125. The method of any one of embodiments 111-124, comprising culturing the host cell in a medium that facilitates post-transcriptional processing of, for example, TREM.

126. The method of any one of embodiments 111-125, comprising culturing the host cell under certain conditions, such as a medium that promotes overexpression or overactivation of enzymes involved in post-transcriptional processing, for example, under conditions that promote:

a) removal of the 5' leader sequence, for example by rnase P;

b) 3' tail exonuclease activity, e.g., rnase II, PNP, rnase PH, or rnase T activity;

c) adding CCA at the 3' end, e.g., by nucleotidyl transferase;

d) intron splicing, for example by one or more (e.g., all) of: a splicing endonuclease, cyclic phosphodiesterase, adenylyltransferase, ligase, or 2' phosphotransferase;

e) modifying, e.g., by modifying an enzyme, e.g., an enzyme having one or more of the following enzymatic activities:

(i) adenosine A₃₄To inosine I₃₄Deamination;

(ii) adenosine m¹A₅₈Methylation of (a);

(iii) proceed to ncm⁵Um₃₄ or ncm⁵s²U₃₄Modifying;

(iv) carry out ct⁶A modification; isoamylation of i⁶A₃₇Modifying; a. the₃₇To i⁶A₃₇Modifying; or

(v) The modifications listed in table 2 were performed; or

f) A synthetase involved in amino acid loading.

127. The method of any one of embodiments 111-126, comprising culturing the host cell in a medium having excess nutrients, e.g., not nutrient limited.

128. The method of any one of embodiments 111-127 comprising culturing the host cell in a medium that promotes expression of Mck1 and/or Kns1, e.g., increases Mck1 and/or Kns1 expression and/or activity.

129. The method of any one of embodiments 111-128, wherein the host cell has increased expression and/or activity of Trm 1.

130. The method of any one of embodiments 111-129, wherein the host cell has reduced activity of Maf1, e.g., by phosphorylation of serine at position 45 of Maf1, e.g., Maf 1.

131. The method of example 130, wherein a decrease in Maf1 activity results in an increase in TREM production.

132. The method of embodiment 130 or 131, wherein the activity of Maf1 can be reduced by: introduction of a pseudo-phosphorylated Maf1 mutant e.g., a mutant having a serine to aspartate mutation at position 45 (S45D); or over-activating CK2/TORC1, for example, which phosphorylates Maf 1.

133. A reaction mixture comprising TREM and a reagent, such as a capture reagent or a separation reagent.

134. A bioreactor comprising a plurality of mammalian host cells described herein comprising exogenous DNA or RNA encoding TREM.

135. The bioreactor of embodiment 134, further comprising a bioreactor

(i) Which comprises at least 1x10⁷、1x 10⁸、1x 10⁹、1x 10¹⁰、1x 10¹¹、1x 10¹²、1x 10¹³Or 1x10¹⁴A host cell;

(ii) comprises 100mL to 100L of a medium, e.g., at least 100mL, 250mL, 500mL, 750mL, 1 liter, 2 liters, 3 liters, 4 liters, 5 liters, 6 liters, 7 liters, 8 liters, 9 liters, 10 liters, 15, 20 liters, 25 liters, 30 liters, 40 liters, 50 liters, 60 liters, 70 liters, 80 liters, 90 liters, or 100 liters of a medium;

(iii) wherein the bioreactor is selected from the group consisting of a continuous flow bioreactor, a batch process bioreactor, a perfusion bioreactor and a fed-batch bioreactor; or

(iv) Wherein the bioreactor is maintained under conditions sufficient to express the TREM.

136. A master cell bank comprising host cells, e.g., as described herein.

137. The master cell library of embodiment 136, wherein the master cell library comprises at least 1x10⁷、1x 10⁸、1x 10⁹、1x 10¹⁰、1x 10¹¹、1x 10¹²、1x 10¹³、1x 10¹⁴、1x 10¹⁵、1x 10²⁰、1x 10²⁵Or 1x10³⁰A host cell.

138. A method of assessing a composition of TREM, e.g., GMP-grade TREM (i.e., a TREM that conforms to cGMP and/or is prepared according to similar requirements), the method comprising obtaining a value for one or more of the following characteristics of the purified TREM composition:

(i) purity of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%;

(iv) DNA, e.g., host cell DNA, of less than 1ng/ml, 5ng/ml, 10ng/ml, 15ng/ml, 20ng/ml, 25ng/ml, 30ng/ml, 35ng/ml, 40ng/ml, 50ng/ml, 60ng/ml, 70ng/ml, 80ng/ml, 90ng/ml or 100 ng/ml;

(v) less than 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% TREM fragments relative to full-length TREM;

(vi) low or absent endotoxin levels, e.g., a negative result as measured by a Limulus Amoebocyte Lysate (LAL) test;

(vii) in vitro translation activity, e.g., as measured by the assay described in example 15;

(ix) Aseptically, e.g., the composition or formulation supports the growth of less than 100 viable microorganisms when tested under aseptic conditions, the composition or formulation meets the criteria of USP <71>, and/or the composition or formulation meets the criteria of USP <85> as described in cGMP guidelines for aseptically processed manufactured sterile pharmaceutical products; or

(x) Viral contamination, e.g., the composition or formulation is not present or no level of viral contamination is detected.

139. The method of any one of examples 1-66, the composition or pharmaceutical composition of any one of examples 67-82, the method of any one of examples 83-109, the cell of any one of examples 110-132, the reaction mixture of example 133, the bioreactor of example 134 or 135, the master cell bank of example 136 or 137, or the method of assessing of example 138, wherein the TREM is encoded by or expressed by a nucleic acid sequence comprising:

(i) a control region sequence;

(ii) a sequence encoding a modified TREM;

(iii) a sequence encoding more than one TREM; or

(iv)tRNA^MetSequences other than the sequence.

140. The method, composition, or pharmaceutical composition, cell, reaction mixture, bioreactor, or master cell bank of example 74, wherein the nucleic acid sequence comprises a promoter sequence.

141. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of embodiment 139 or 140 wherein the nucleic acid sequence comprises a promoter sequence comprising an RNA polymerase III (Pol III) recognition site, e.g., a Pol III binding site, e.g., a U6 promoter sequence or fragment thereof.

142. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank as described in any one of examples 139-141, wherein the nucleic acid sequence comprises a promoter sequence comprising a mutation, such as a promoter up-regulation mutation, for example a mutation that increases transcription initiation, for example a mutation that increases TFIIIB binding.

143. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 139-142, wherein the nucleic acid sequence comprises a promoter sequence that increases Pol III binding and results in increased tRNA production, e.g., TREM production.

144. The method of any one of examples 1-66 or 139-143, the composition or the pharmaceutical composition of any one of examples 67-82 or 139-143, the method of any one of examples 83-109 or 139-143, the cell of any one of examples 110-132 or 139-143, the reaction mixture of example 133 or 139-143, the bioreactor of example 134-135 or 139-143, the main cell bank of example 136-137 or 139-143, or the method of assessing of example 138 or 139-143, wherein the TREM enhances:

(a) Stability of the product, e.g. of the protein, and/or

(b) Ribosome occupancy of the product.

145. The method of any one of examples 1-66 or 139-144, the composition or the pharmaceutical composition of any one of examples 67-82 or 139-144, the method of any one of examples 83-109 or 139-144, the cell of any one of examples 110-132 or 139-144, the reaction mixture of example 133 or 139-144, the bioreactor of example 134-135 or 139-144, the main cell bank of example 136-137 or 139-144, or the method of assessment of example 138 or 139-144, wherein the TREM:

regulating ribosome occupancy;

regulating protein translation or stability;

modulating mRNA stability;

modulating protein folding or structure;

regulating protein transduction or compartmentalization;

(ii) modulation of codon usage;

modulating cell fate; or

Modulating signaling pathways, such as cellular signaling pathways.

146. The method of any one of examples 1-66 or 139-144, the composition or the pharmaceutical composition of any one of examples 67-82 or 139-144, the method of any one of examples 83-109 or 139-144, the cell of any one of examples 110-132 or 139-144, the reaction mixture of example 133 or 139-144, the bioreactor of example 134-135 or 139-144, the primary cell bank of example 136-137 or 139-144, or the method of assessment of example 138 or 139-144, wherein the TREM comprises post-transcriptional modifications from Table 2.

147. The preparation method according to any one of examples 1-66 or 139-146, the composition or the pharmaceutical composition according to any one of examples 67-82 or 139-146, the method according to any one of examples 83-109 or 139-146, the cell according to any one of examples 110-132 or 139-146, the reaction mixture according to example 133 or 139-146, the bioreactor according to example 134-135 or 139-146, the main cell bank according to example 136-137 or 139-146, or the evaluation method according to example 138 or 139-146, wherein the TREM comprises a homologous adaptor function, and wherein during initiation or elongation of a peptide chain, the TREM mediates acceptance and incorporation of an amino acid naturally associated with the anticodon of the TREM.

148. The method of any one of examples 1-66 or 139-147, the composition or pharmaceutical composition of any one of examples 67-82 or 139-147, the method of any one of examples 83-109 or 139-147, the cell of any one of examples 110-132 or 139-147, the reaction mixture of example 133 or 139-147, the bioreactor of example 134-135 or 139-147, the primary cell library of example 136-137 or 139-147, or the method of assessment of example 138 or 139-147, wherein the TREM comprises a non-homologous adaptor function, and wherein the TREM mediates the acceptance and incorporation of amino acids other than the amino acid naturally associated with the anticodon of the TREM during the initiation or extension of the peptide chain, e.g., non-homologous amino acids, and the non-homologous amino acid residues are, e.g., desired residues, e.g., residues that do not mediate a disorder or unwanted trait, e.g., wild-type residues.

149. The method of any one of examples 1-66 or 139- -

Specifying a first amino acid residue, e.g., an undesired or undesirable codon, e.g., a codon associated with a disorder or undesired trait, e.g., a mutant codon, and

the TREM mediates incorporation of a second amino acid residue, e.g., a desired codon, e.g., an amino acid not associated with a disorder or unwanted trait, e.g., a wild-type amino acid.

150. The method of any one of examples 1-66 or 139-149, the composition or pharmaceutical composition of any one of examples 67-75, 79-82 or 139-149, the method of any one of examples 83-109 or 139-149, the cell of any one of examples 110-132 or 139-149, the reaction mixture of examples 133 or 139-149, the bioreactor of examples 134-135 or 139-149, the main cell bank of examples 136-137 or 139-149, or the evaluation method of examples 138 or 139-149, wherein the TREM comprises an RNA sequence that is at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%) identical to the RNA sequence of the naturally-occurring tRNA.

151. The method of any one of examples 1-66 or 139-150, the composition or the pharmaceutical composition of any one of examples 67-75, 79-82 or 139-150, the method of any one of examples 83-109 or 139-150, the cell of any one of examples 110-132 or 139-150, the reaction mixture of example 133 or 139-150, the bioreactor of example 134-135 or 139-150, the host cell bank of example 136-137 or 139-150, or the evaluation method of example 138 or 139-150, wherein the TREM comprises at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 97%, at least 98% >, or at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 97%, at least 98% >, or at least one of the RNA encoded by the DNA sequences listed in Table 1, At least 99%) identical RNA sequences, or fragments or functional fragments thereof.

152. The method of any one of examples 1-66 or 139-151, the composition or the pharmaceutical composition of any one of examples 67-75, 79-82 or 139-151, the method of any one of examples 83-109 or 139-151, the cell of any one of examples 110-132 or 139-151, the reaction mixture of example 133 or 139-151, the bioreactor of example 134-135 or 139-151, the main cell library of example 136-137 or 139-151, or the evaluation method of example 138 or 139-151, wherein the TREM comprises:

An RNA sequence encoded by a DNA sequence listed in table 1, or a fragment thereof.

153. The method of any one of examples 1-66 or 139-152, the composition or the pharmaceutical composition of any one of examples 67-75, 79-82 or 139-152, the method of any one of examples 83-109 or 139-152, the cell of any one of examples 110-132 or 139-152, the reaction mixture of example 133 or 139-152, the bioreactor of example 134-135 or 139-152, the main cell library of example 136-137 or 139-152, or the evaluation method of example 138 or 139-152, wherein the TREM comprises:

an RNA sequence at least XX% identical to an RNA sequence encoded by a DNA sequence listed in table 1, or a fragment thereof, wherein XX is selected from 80, 85, 90, 95, 96, 97, 98 or 99.

154. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 153 wherein XX is 80.

155. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 153 wherein XX is 85.

156. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 153 wherein XX is 90.

157. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 153 wherein XX is 95.

158. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 153 wherein XX is 97.

159. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 153 wherein XX is 98.

160. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 153 wherein XX is 99.

161. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank as described in any one of examples 153-160, wherein the DNA sequence is SEQ ID NO 1 or a fragment thereof, or SEQ ID NO 2 or a fragment thereof, or SEQ ID NO 3 or a fragment thereof, or SEQ ID NO 4 or a fragment thereof, or SEQ ID NO 5 or a fragment thereof, or SEQ ID NO 6 or a fragment thereof, or SEQ ID NO 7 or a fragment thereof, or SEQ ID NO 8 or a fragment thereof, or SEQ ID NO 9 or a fragment thereof, or SEQ ID NO 10 or a fragment thereof, or SEQ ID NO 11 or a fragment thereof, or SEQ ID NO 12 or a fragment thereof, or SEQ ID NO 13 or a fragment thereof, or SEQ ID NO 14 or a fragment thereof, or SEQ ID NO 15 or a fragment thereof, or SEQ ID NO 16 or a fragment thereof, Or SEQ ID NO 17 or a fragment thereof, or SEQ ID NO 18 or a fragment thereof, or SEQ ID NO 19 or a fragment thereof, or SEQ ID NO 20 or a fragment thereof, or SEQ ID NO 21 or a fragment thereof, or SEQ ID NO 22 or a fragment thereof, or SEQ ID NO 23 or a fragment thereof, or SEQ ID NO 24 or a fragment thereof, or SEQ ID NO 25 or a fragment thereof, or SEQ ID NO 26 or a fragment thereof, or SEQ ID NO 27 or a fragment thereof, or SEQ ID NO 28 or a fragment thereof, or SEQ ID NO 29 or a fragment thereof, or SEQ ID NO 30 or a fragment thereof, or SEQ ID NO 31 or a fragment thereof, or SEQ ID NO 32 or a fragment thereof, or SEQ ID NO 33 or a fragment thereof, or SEQ ID NO 34 or a fragment thereof, or SEQ ID NO 35 or a fragment thereof, or SEQ ID NO 36 or a fragment thereof, Or SEQ ID NO 37 or a fragment thereof, or SEQ ID NO 38 or a fragment thereof, or SEQ ID NO 39 or a fragment thereof, or SEQ ID NO 40 or a fragment thereof, or SEQ ID NO 41 or a fragment thereof, or SEQ ID NO 42 or a fragment thereof, or SEQ ID NO 43 or a fragment thereof, or SEQ ID NO 44 or a fragment thereof, or SEQ ID NO 45 or a fragment thereof, or SEQ ID NO 46 or a fragment thereof, or SEQ ID NO 47 or a fragment thereof, or SEQ ID NO 48 or a fragment thereof, or SEQ ID NO 49 or a fragment thereof, or SEQ ID NO 50 or a fragment thereof, or SEQ ID NO 51 or a fragment thereof, or SEQ ID NO 52 or a fragment thereof, or SEQ ID NO 53 or a fragment thereof, or SEQ ID NO 54 or a fragment thereof, or SEQ ID NO 55 or a fragment thereof, or SEQ ID NO 56 or a fragment thereof, Or SEQ ID NO 57 or a fragment thereof, or SEQ ID NO 58 or a fragment thereof, or SEQ ID NO 59 or a fragment thereof, or SEQ ID NO 60 or a fragment thereof, or SEQ ID NO 61 or a fragment thereof, or SEQ ID NO 62 or a fragment thereof, or SEQ ID NO 63 or a fragment thereof, or SEQ ID NO 64 or a fragment thereof, or SEQ ID NO 65 or a fragment thereof, or SEQ ID NO 66 or a fragment thereof, or SEQ ID NO 67 or a fragment thereof, or SEQ ID NO 68 or a fragment thereof, or SEQ ID NO 69 or a fragment thereof, or SEQ ID NO 70 or a fragment thereof, or SEQ ID NO 71 or a fragment thereof, or SEQ ID NO 72 or a fragment thereof, or SEQ ID NO 73 or a fragment thereof, or SEQ ID NO 74 or a fragment thereof, or SEQ ID NO 75 or a fragment thereof, or SEQ ID NO 76 or a fragment thereof, Or SEQ ID NO 77 or a fragment thereof, or SEQ ID NO 78 or a fragment thereof, or SEQ ID NO 79 or a fragment thereof, or SEQ ID NO 80 or a fragment thereof, or SEQ ID NO 81 or a fragment thereof, or SEQ ID NO 82 or a fragment thereof, or SEQ ID NO 83 or a fragment thereof, or SEQ ID NO 84 or a fragment thereof, or SEQ ID NO 85 or a fragment thereof, or SEQ ID NO 86 or a fragment thereof, or SEQ ID NO 87 or a fragment thereof, or SEQ ID NO 88 or a fragment thereof, or SEQ ID NO 89 or a fragment thereof, or SEQ ID NO 90 or a fragment thereof, or SEQ ID NO 91 or a fragment thereof, or SEQ ID NO 92 or a fragment thereof, or SEQ ID NO 93 or a fragment thereof, or SEQ ID NO 94 or a fragment thereof, or SEQ ID NO 95 or a fragment thereof, or SEQ ID NO 96 or a fragment thereof, Or SEQ ID NO 97 or a fragment thereof, or SEQ ID NO 98 or a fragment thereof, or SEQ ID NO 99 or a fragment thereof, or SEQ ID NO 100 or a fragment thereof, or SEQ ID NO 101 or a fragment thereof, or SEQ ID NO 102 or a fragment thereof, or SEQ ID NO 103 or a fragment thereof, or SEQ ID NO 104 or a fragment thereof, or SEQ ID NO 105 or a fragment thereof, or SEQ ID NO 106 or a fragment thereof, or SEQ ID NO 107 or a fragment thereof, or SEQ ID NO 108 or a fragment thereof, or SEQ ID NO 109 or a fragment thereof, or SEQ ID NO 110 or a fragment thereof, or SEQ ID NO 111 or a fragment thereof, or SEQ ID NO 112 or a fragment thereof, or SEQ ID NO 113 or a fragment thereof, or SEQ ID NO 114 or a fragment thereof, or SEQ ID NO 115 or a fragment thereof, or SEQ ID NO 116 or a fragment thereof, Or SEQ ID NO 117 or a fragment thereof, or SEQ ID NO 118 or a fragment thereof, or SEQ ID NO 119 or a fragment thereof, or SEQ ID NO 120 or a fragment thereof, or SEQ ID NO 121 or a fragment thereof, or SEQ ID NO 122 or a fragment thereof, or SEQ ID NO 123 or a fragment thereof, or SEQ ID NO 124 or a fragment thereof, or SEQ ID NO 125 or a fragment thereof, or SEQ ID NO 126 or a fragment thereof, or SEQ ID NO 127 or a fragment thereof, or SEQ ID NO 128 or a fragment thereof, or SEQ ID NO 129 or a fragment thereof, or SEQ ID NO 130 or a fragment thereof, or SEQ ID NO 131 or a fragment thereof, or SEQ ID NO 132 or a fragment thereof, or SEQ ID NO 133 or a fragment thereof, or SEQ ID NO 134 or a fragment thereof, or SEQ ID NO 135 or a fragment thereof, or SEQ ID NO 136 or a fragment thereof, Or SEQ ID NO 137 or a fragment thereof, or SEQ ID NO 138 or a fragment thereof, or SEQ ID NO 139 or a fragment thereof, or SEQ ID NO 140 or a fragment thereof, or SEQ ID NO 141 or a fragment thereof, or SEQ ID NO 142 or a fragment thereof, or SEQ ID NO 143 or a fragment thereof, or SEQ ID NO 144 or a fragment thereof, or SEQ ID NO 145 or a fragment thereof, or SEQ ID NO 146 or a fragment thereof, or SEQ ID NO 147 or a fragment thereof, or SEQ ID NO 148 or a fragment thereof, or SEQ ID NO 149 or a fragment thereof, or SEQ ID NO 150 or a fragment thereof, or SEQ ID NO 151 or a fragment thereof, or SEQ ID NO 152 or a fragment thereof, or SEQ ID NO 153 or a fragment thereof, or SEQ ID NO 154 or a fragment thereof, or SEQ ID NO 155 or a fragment thereof, or SEQ ID NO 156 or a fragment thereof, Or SEQ ID NO 157 or a fragment thereof, or SEQ ID NO 158 or a fragment thereof, or SEQ ID NO 159 or a fragment thereof, or SEQ ID NO 160 or a fragment thereof, or SEQ ID NO 161 or a fragment thereof, or SEQ ID NO 162 or a fragment thereof, or SEQ ID NO 163 or a fragment thereof, or SEQ ID NO 164 or a fragment thereof, or SEQ ID NO 165 or a fragment thereof, or SEQ ID NO 166 or a fragment thereof, or SEQ ID NO 167 or a fragment thereof, or SEQ ID NO 168 or a fragment thereof, or SEQ ID NO 169 or a fragment thereof, or SEQ ID NO 170 or a fragment thereof, or SEQ ID NO 171 or a fragment thereof, or SEQ ID NO 172 or a fragment thereof, or SEQ ID NO 173 or a fragment thereof, or SEQ ID NO 174 or a fragment thereof, or SEQ ID NO 175 or a fragment thereof, or SEQ ID NO 176 or a fragment thereof, Or SEQ ID NO 177 or a fragment thereof, or SEQ ID NO 178 or a fragment thereof, or SEQ ID NO 179 or a fragment thereof, or SEQ ID NO 180 or a fragment thereof, or SEQ ID NO 181 or a fragment thereof, or SEQ ID NO 182 or a fragment thereof, or SEQ ID NO 183 or a fragment thereof, or SEQ ID NO 184 or a fragment thereof, or SEQ ID NO 185 or a fragment thereof, or SEQ ID NO 186 or a fragment thereof, or SEQ ID NO 187 or a fragment thereof, or SEQ ID NO 188 or a fragment thereof, or SEQ ID NO 189 or a fragment thereof, or SEQ ID NO 190 or a fragment thereof, or SEQ ID NO 191 or a fragment thereof, or SEQ ID NO 192 or a fragment thereof, or SEQ ID NO 193 or a fragment thereof, or SEQ ID NO 194 or a fragment thereof, or SEQ ID NO 195 or a fragment thereof, or SEQ ID NO 196 or a fragment thereof, Or SEQ ID NO 197 or a fragment thereof, or SEQ ID NO 198 or a fragment thereof, or SEQ ID NO 199 or a fragment thereof, or SEQ ID NO 200 or a fragment thereof, or SEQ ID NO 201 or a fragment thereof, or SEQ ID NO 202 or a fragment thereof, or SEQ ID NO 203 or a fragment thereof, or SEQ ID NO 204 or a fragment thereof, or SEQ ID NO 205 or a fragment thereof, or SEQ ID NO 206 or a fragment thereof, or SEQ ID NO 207 or a fragment thereof, or SEQ ID NO 208 or a fragment thereof, or SEQ ID NO 209 or a fragment thereof, or SEQ ID NO 210 or a fragment thereof, or SEQ ID NO 211 or a fragment thereof, or SEQ ID NO 212 or a fragment thereof, or SEQ ID NO 213 or a fragment thereof, or SEQ ID NO 214 or a fragment thereof, or SEQ ID NO 215 or a fragment thereof, or SEQ ID NO 216 or a fragment thereof, Or SEQ ID NO 217 or a fragment thereof, or SEQ ID NO 218 or a fragment thereof, or SEQ ID NO 219 or a fragment thereof, or SEQ ID NO 220 or a fragment thereof, or SEQ ID NO 221 or a fragment thereof, or SEQ ID NO 222 or a fragment thereof, or SEQ ID NO 223 or a fragment thereof, or SEQ ID NO 224 or a fragment thereof, or SEQ ID NO 225 or a fragment thereof, or SEQ ID NO 226 or a fragment thereof, or SEQ ID NO 227 or a fragment thereof, or SEQ ID NO 228 or a fragment thereof, or SEQ ID NO 229 or a fragment thereof, or SEQ ID NO 230 or a fragment thereof, or SEQ ID NO 231 or a fragment thereof, or SEQ ID NO 232 or a fragment thereof, or SEQ ID NO 233 or a fragment thereof, or SEQ ID NO 234 or a fragment thereof, or SEQ ID NO 235 or a fragment thereof, or SEQ ID NO 236 or a fragment thereof, Or SEQ ID NO 237 or a fragment thereof, or SEQ ID NO 238 or a fragment thereof, or SEQ ID NO 239 or a fragment thereof, or SEQ ID NO 240 or a fragment thereof, or SEQ ID NO 241 or a fragment thereof, or SEQ ID NO 242 or a fragment thereof, or SEQ ID NO 243 or a fragment thereof, or SEQ ID NO 244 or a fragment thereof, or SEQ ID NO 245 or a fragment thereof, or SEQ ID NO 246 or a fragment thereof, or SEQ ID NO 247 or a fragment thereof, or SEQ ID NO 248 or a fragment thereof, or SEQ ID NO 249 or a fragment thereof, or SEQ ID NO 250 or a fragment thereof, or SEQ ID NO 251 or a fragment thereof, or SEQ ID NO 252 or a fragment thereof, or SEQ ID NO 253 or a fragment thereof, or SEQ ID NO 254 or a fragment thereof, or SEQ ID NO 255 or a fragment thereof, or SEQ ID NO 256 or a fragment thereof, Or SEQ ID NO 257 or a fragment thereof, or SEQ ID NO 258 or a fragment thereof, or SEQ ID NO 259 or a fragment thereof, or SEQ ID NO 260 or a fragment thereof, or SEQ ID NO 261 or a fragment thereof, or SEQ ID NO 262 or a fragment thereof, or SEQ ID NO 263 or a fragment thereof, or SEQ ID NO 264 or a fragment thereof, or SEQ ID NO 265 or a fragment thereof, or SEQ ID NO 266 or a fragment thereof, or SEQ ID NO 267 or a fragment thereof, or SEQ ID NO 268 or a fragment thereof, or SEQ ID NO 269 or a fragment thereof, or SEQ ID NO 270 or a fragment thereof, or SEQ ID NO 271 or a fragment thereof, or SEQ ID NO 272 or a fragment thereof, or SEQ ID NO 273 or a fragment thereof, or SEQ ID NO 274 or a fragment thereof, or SEQ ID NO 275 or a fragment thereof, or SEQ ID NO 276 or a fragment thereof, Or SEQ ID NO 277 or a fragment thereof, or SEQ ID NO 278 or a fragment thereof, or SEQ ID NO 279 or a fragment thereof, or SEQ ID NO 280 or a fragment thereof, or SEQ ID NO 281 or a fragment thereof, or SEQ ID NO 282 or a fragment thereof, or SEQ ID NO 283 or a fragment thereof, or SEQ ID NO 284 or a fragment thereof, or SEQ ID NO 285 or a fragment thereof, or SEQ ID NO 286 or a fragment thereof, or SEQ ID NO 287 or a fragment thereof, or SEQ ID NO 288 or a fragment thereof, or SEQ ID NO 290 or a fragment thereof, or SEQ ID NO 291 or a fragment thereof, or SEQ ID NO 292 or a fragment thereof, or SEQ ID NO 293 or a fragment thereof, or SEQ ID NO 294 or a fragment thereof, or SEQ ID NO 295 or a fragment thereof, or SEQ ID NO 296 or a fragment thereof, Or SEQ ID NO 297 or a fragment thereof, or SEQ ID NO 298 or a fragment thereof, or SEQ ID NO 299 or a fragment thereof, or SEQ ID NO 300 or a fragment thereof, or SEQ ID NO 301 or a fragment thereof, or SEQ ID NO 302 or a fragment thereof, or SEQ ID NO 303 or a fragment thereof, or SEQ ID NO 304 or a fragment thereof, or SEQ ID NO 305 or a fragment thereof, or SEQ ID NO 306 or a fragment thereof, or SEQ ID NO 307 or a fragment thereof, or SEQ ID NO 308 or a fragment thereof, or SEQ ID NO 309 or a fragment thereof, or SEQ ID NO 310 or a fragment thereof, or SEQ ID NO 311 or a fragment thereof, or SEQ ID NO 312 or a fragment thereof, or SEQ ID NO 313 or a fragment thereof, or SEQ ID NO 314 or a fragment thereof, or SEQ ID NO 315 or a fragment thereof, or SEQ ID NO 316 or a fragment thereof, Or SEQ ID NO 317 or a fragment thereof, or SEQ ID NO 318 or a fragment thereof, or SEQ ID NO 319 or a fragment thereof, or SEQ ID NO 320 or a fragment thereof, or SEQ ID NO 321 or a fragment thereof, or SEQ ID NO 322 or a fragment thereof, or SEQ ID NO 323 or a fragment thereof, or SEQ ID NO 324 or a fragment thereof, or SEQ ID NO 325 or a fragment thereof, or SEQ ID NO 326 or a fragment thereof, or SEQ ID NO 327 or a fragment thereof, or SEQ ID NO 328 or a fragment thereof, or SEQ ID NO 329 or a fragment thereof, or SEQ ID NO 330 or a fragment thereof, or SEQ ID NO 331 or a fragment thereof, or SEQ ID NO 332 or a fragment thereof, or SEQ ID NO 333 or a fragment thereof, or SEQ ID NO 334 or a fragment thereof, or SEQ ID NO 335 or a fragment thereof, or SEQ ID NO 336 or a fragment thereof, Or SEQ ID NO 337 or a fragment thereof, or SEQ ID NO 338 or a fragment thereof, or SEQ ID NO 339 or a fragment thereof, or SEQ ID NO 340 or a fragment thereof, or SEQ ID NO 341 or a fragment thereof, or SEQ ID NO 342 or a fragment thereof, or SEQ ID NO 343 or a fragment thereof, or SEQ ID NO 344 or a fragment thereof, or SEQ ID NO 345 or a fragment thereof, or SEQ ID NO 346 or a fragment thereof, or SEQ ID NO 347 or a fragment thereof, or SEQ ID NO 348 or a fragment thereof, or SEQ ID NO 349 or a fragment thereof, or SEQ ID NO 350 or a fragment thereof, or SEQ ID NO 351 or a fragment thereof, or SEQ ID NO 352 or a fragment thereof, or SEQ ID NO 353 or a fragment thereof, or SEQ ID NO 354 or a fragment thereof, or SEQ ID NO 355 or a fragment thereof, or SEQ ID NO 356 or a fragment thereof, Or SEQ ID NO. 357 or a fragment thereof, or SEQ ID NO. 358 or a fragment thereof, or SEQ ID NO. 359 or a fragment thereof, or SEQ ID NO. 360 or a fragment thereof, or SEQ ID NO. 361 or a fragment thereof, or SEQ ID NO. 362 or a fragment thereof, or SEQ ID NO. 363 or a fragment thereof, or SEQ ID NO. 364 or a fragment thereof, or SEQ ID NO. 365 or a fragment thereof, or SEQ ID NO. 366 or a fragment thereof, or SEQ ID NO. 367 or a fragment thereof, or SEQ ID NO. 368 or a fragment thereof, or SEQ ID NO. 369 or a fragment thereof, or SEQ ID NO. 370 or a fragment thereof, or SEQ ID NO. 371 or a fragment thereof, SEQ ID NO. 372 or a fragment thereof, or SEQ ID NO. 373 or a fragment thereof, or SEQ ID NO. 374 or a fragment thereof, or SEQ ID NO. 375 or a fragment thereof, or SEQ ID NO. 376 or a fragment thereof, Or SEQ ID NO 377 or fragment thereof, or SEQ ID NO 378 or fragment thereof, or SEQ ID NO 379 or fragment thereof, or SEQ ID NO 380 or fragment thereof, or SEQ ID NO 381 or fragment thereof, or SEQ ID NO 382 or fragment thereof, or SEQ ID NO 383 or fragment thereof, or SEQ ID NO 384 or fragment thereof, or SEQ ID NO 385 or fragment thereof, or SEQ ID NO 386 or fragment thereof, or SEQ ID NO 387 or fragment thereof, or SEQ ID NO 388 or fragment thereof, or SEQ ID NO 389 or fragment thereof, or SEQ ID NO 390 or fragment thereof, or SEQ ID NO 391 or fragment thereof, or SEQ ID NO 392 or fragment thereof, or SEQ ID NO 393 or fragment thereof, or SEQ ID NO 394 or fragment thereof, or SEQ ID NO 395 or fragment thereof, or SEQ ID NO 396 or fragment thereof, Or SEQ ID NO:397 or a fragment thereof, or SEQ ID NO:398 or a fragment thereof, or SEQ ID NO:399 or a fragment thereof, or SEQ ID NO:400 or a fragment thereof, or SEQ ID NO:401 or a fragment thereof, or SEQ ID NO:402 or a fragment thereof, or SEQ ID NO:403 or a fragment thereof, or SEQ ID NO:404 or a fragment thereof, or SEQ ID NO:405 or a fragment thereof, or SEQ ID NO:406 or a fragment thereof, or SEQ ID NO:407 or a fragment thereof, or SEQ ID NO:408 or a fragment thereof, or SEQ ID NO:409 or a fragment thereof, or SEQ ID NO:410 or a fragment thereof, or SEQ ID NO:411 or a fragment thereof, or SEQ ID NO:412 or a fragment thereof, or SEQ ID NO:413 or a fragment thereof, or SEQ ID NO:414 or a fragment thereof, or SEQ ID NO:415 or a fragment thereof, or SEQ ID NO:416 or a fragment thereof, Or SEQ ID NO 417 or a fragment thereof, or SEQ ID NO 418 or a fragment thereof, or SEQ ID NO 419 or a fragment thereof, or SEQ ID NO 420 or a fragment thereof, or SEQ ID NO 421 or a fragment thereof, or SEQ ID NO 422 or a fragment thereof, or SEQ ID NO 423 or a fragment thereof, or SEQ ID NO 424 or a fragment thereof, or SEQ ID NO 425 or a fragment thereof, or SEQ ID NO 426 or a fragment thereof, or SEQ ID NO 427 or a fragment thereof, or SEQ ID NO 428 or a fragment thereof, or SEQ ID NO 429 or a fragment thereof, or SEQ ID NO 430 or a fragment thereof, or SEQ ID NO 431 or a fragment thereof, or SEQ ID NO 432 or a fragment thereof, or SEQ ID NO 433 or a fragment thereof, or SEQ ID NO 434 or a fragment thereof, or SEQ ID NO 435 or a fragment thereof, or SEQ ID NO 436 or a fragment thereof, Or SEQ ID NO 437 or a fragment thereof, or SEQ ID NO 438 or a fragment thereof, or SEQ ID NO 439 or a fragment thereof, or SEQ ID NO 440 or a fragment thereof, or SEQ ID NO 441 or a fragment thereof, or SEQ ID NO 442 or a fragment thereof, or SEQ ID NO 443 or a fragment thereof, or SEQ ID NO 444 or a fragment thereof, or SEQ ID NO 445 or a fragment thereof, or SEQ ID NO 446 or a fragment thereof, or SEQ ID NO 447 or a fragment thereof, or SEQ ID NO 448 or a fragment thereof, or SEQ ID NO 449 or a fragment thereof, or SEQ ID NO 450 or a fragment thereof, or SEQ ID NO 451 or a fragment thereof,

Optionally, wherein fragments include one or more, but not all, of: linker 1 region, AStD stem region; a linker 2 region; stem-loop regions, e.g., the D-arm region; a linker 3 region; stem-loop regions, e.g., AC arm regions; a variable region; stem-loop regions, e.g., T-arm regions; and linker 4 regions, e.g., such as those described herein.

162. The composition or pharmaceutical composition of any one of examples 76 to 82, the method of any one of examples 94 to 109, or the cell of any one of claims 110-132, wherein ZZZ represents any one of the following amino acids: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine or valine.

163. The method of any one of examples 1-66 or 139-161, the composition or the pharmaceutical composition of any one of examples 67-75, 79-82 or 139-161, the method of any one of examples 83-109 or 139-161, the cell of any one of examples 110-132 or 139-161, the reaction mixture of example 133 or 139-161, the bioreactor of example 134-135 or 139-161, the host cell bank of example 136-137 or 139-161, or the method of assessing of example 138 or 139-161, wherein the TREM comprises a property selected from the group consisting of (e.g., in the cell bank having the structure R-66, 79-82 or 139-161), the TREM comprises a property selected from the group consisting of ₀-R_1-R₂-R₃-R₄-R₅-R₆-R₇-R₈-R₉-R₁₀-R₁₁-R₁₂-R₁₃-R₁₄-R₁₅-R₁₆-R₁₇-R₁₈-R₁₉-R₂₀-R₂₁-R₂₂-R₂₃-R₂₄-R₂₅-R₂₆-R₂₇-R₂₈-R₂₉-R₃₀-R₃₁-R₃₂-R₃₃-R₃₄-R₃₅-R₃₆-R₃₇-R₃₈-R₃₉-R₄₀-R₄₁-R₄₂-R₄₃-R₄₄-R₄₅-R₄₆-[R₄₇]_x-R₄₈-R₄₉-R₅₀-R₅₁-R₅₂-R₅₃-R₅₄-R₅₅-R₅₆-R₅₇-R₅₈-R₅₉-R₆₀-R₆₁-R₆₂-R₆₃-R₆₄-R₆₅-R₆₆-R₆₇-R₆₈-R₆₉-R₇₀-R₇₁-R₇₂Wherein R is a ribonucleotide residue):

a) under physiological conditions, the residue R₀Forming a linker region, e.g., linker 1 region;

b) residue R under physiological conditions₁-R₂-R₃-R₄-R₅-R₆-R₇And a residue R₆₅-R₆₆-R₆₇-R₆₈-R₆₉-R₇₀-R₇₁Forming a stem region, e.g., an AStD stem region;

c) under physiological conditions, the residue R₈-R₉Forming a linker region, e.g., linker 2 region;

d) residue-R under physiological conditions₁₀-R₁₁-R₁₂-R₁₃-R₁₄R₁₅-R₁₆-R₁₇-R₁₈-R₁₉-R₂₀-R₂₁-R₂₂-R₂₃-R₂₄-R₂₅-R₂₆-R₂₇-R₂₈Forming a stem-loop region, e.g., a D-arm region;

e) residue-R under physiological conditions₂₉Forming a linker region, e.g., linker 3 region;

f) residue-R under physiological conditions₃₀-R₃₁-R₃₂-R₃₃-R₃₄-R₃₅-R₃₆-R₃₇-R₃₈-R₃₉-R₄₀-R₄₁-R₄₂-R₄₃-R₄₄-R₄₅-R₄₆Forming a stem-loop region, e.g., an AC arm region;

g) residue- [ R ] under physiological conditions₄₇]_xComprises a variable region;

h) residue-R under physiological conditions₄₈-R₄₉-R₅₀-R₅₁-R₅₂-R₅₃-R₅₄-R₅₅-R₅₆-R₅₇-R₅₈-R₅₉-R₆₀-R₆₁-R₆₂-R₆₃-R₆₄Forming a stem-loop region, e.g., a T-arm region; or

i) Residue R under physiological conditions₇₂A tab region, e.g., tab 4 region, is formed.

164. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of embodiment 163 comprising any one of properties (a) - (i).

165. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of embodiment 163 comprising any two of characteristics (a) - (i).

166. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 163 comprising any three of characteristics (a) - (i).

167. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of embodiment 163 comprising any four of characteristics (a) - (i).

168. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 163 comprising any five of characteristics (a) - (i).

169. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 163 comprising any six of characteristics (a) - (i).

170. The composition or pharmaceutical composition, method, or cell of embodiment 163, comprising any seven of properties (a) - (i).

171. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 163 comprising all of characteristics (a) - (i).

172. The method of any one of examples 1-66 or 139-171, the composition or the pharmaceutical composition of any one of examples 67-82 or 139-171, the method of any one of examples 83-109 or 139-171, the cell of any one of examples 110-132 or 139-171, the reaction mixture of example 133 or 139-171, the bioreactor of example 134-135 or 139-171, the main cell bank of example 136-137 or 139-171, or the method of assessment of example 138 or 139-171, wherein the TREM comprises the consensus sequence provided herein.

173. The preparation method according to any one of examples 1-66 or 139-171, the composition or the pharmaceutical composition according to any one of examples 67-82 or 139-171, the composition according to any one of examples 83-109 or 139-171Method, the cell as defined in any one of examples 110-132 or 139-171, the reaction mixture as defined in example 133 or 139-171, the bioreactor as defined in example 134-135 or 139-171, the master cell library as defined in example 136-137 or 139-171, or the evaluation method as defined in example 138 or 139-171, wherein the TREM comprises the formula I_ZZZWherein_ZZZRepresents any of the twenty amino acids and formula I corresponds to all species.

174. The method of any one of examples 1-66 or 139-171, the composition or the pharmaceutical composition of any one of examples 67-82 or 139-171, the method of any one of examples 83-109 or 139-171, the cell of any one of examples 110-132 or 139-171, the reaction mixture of example 133 or 139-171, the bioreactor of example 134-135 or 139-171, the main cell bank of example 136-137 or 139-171, or the method of assessing of example 138 or 139-171, wherein the TREM comprises the formula II _ZZZWherein_ZZZRepresents any of the twenty amino acids and formula II corresponds to a mammal.

175. The method of any one of examples 1-66 or 139-171, the composition or the pharmaceutical composition of any one of examples 67-82 or 139-171, the method of any one of examples 83-109 or 139-171, the cell of any one of examples 110-132 or 139-171, the reaction mixture of example 133 or 139-171, the bioreactor of example 134-135 or 139-171, the main cell bank of example 136-137 or 139-171, or the method of assessing of example 138 or 139-171, wherein the TREM comprises the formula III_ZZZWherein_ZZZRepresents any of the twenty amino acids and formula III corresponds to human.

176. The method of any one of examples 1-66 or 139-175, the composition or pharmaceutical composition of any one of examples 67-82 or 139-175, the method of any one of examples 83-109 or 139-175, the cell of any one of examples 110-132 or 139-175, the method of any one of examples 110-132 or 139-175, and the method of any one of the methods of examples 67-82 or 139-175The reaction mixture of example 133 or 139- ₄₇And includes a variable region.

177. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 176, wherein the variable region is 1-271 residues in length (e.g., 1-250, 1-225, 1-200, 1-175, 1-150, 1-125, 1-100, 1-75, 1-50, 1-40, 1-30, 1-29, 1-28, 1-27, 1-26, 1-25, 1-24, 1-23, 1-22, 1-21, 1-20, 1-19, 1-18, 1-17, 1-16, 1-15, 1-14, 1-13, 1-12, 1-11, 1-10, 10-271, etc.) 20-271, 30-271, 40-271, 50-271, 60-271, 70-271, 80-271, 100-271, 125-271, 150-271, 175-271, 200-271, 225-271, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 50, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200, 225, 250, or 271 residues).

178. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of embodiment 176 or 177, wherein said variable region comprises any one, all or a combination of adenine, cytosine, guanine or uracil.

179. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of examples 176-178, wherein the variable region comprises a ribonucleic acid (RNA) sequence encoded by a deoxyribonucleic acid (DNA) sequence as disclosed in Table 3, e.g. any one of SEQ ID NO:452-561 as disclosed in Table 3.

180. The method of preparation as described in any of examples 1-66 or 139-minus 179, the composition or pharmaceutical composition as described in any of examples 67-82 or 139-minus 179, the method as described in any of examples 83-109 or 139-minus 179, the cell as described in any of examples 110-132 or 139-minus 179, the reaction mixture as described in example 133 or 139-minus 179, the bioreactor as described in example 134-135 or 139-minus 179, the master cell bank as described in example 136-137 or 139-minus 179, or the method of assessment as described in example 138 or 139-minus 179, wherein the TREM comprises properties from (e.g., one, two, three, four, five, six, seven, eight, nine or all, or any combination thereof):

a) if the TREM, e.g., if the AC stem loop of the TREM, comprises an exogenous insert, the exogenous insert is no more than 5 contiguous ribonucleotide residues in length;

b) If the TREM, e.g., if the AC stem loop of the TREM, comprises an exogenous insert, the remainder of the molecule comprises a non-naturally occurring sequence, e.g., a non-naturally occurring sequence of 1, 2, 3, 4, 5 or more ribonucleotide residues;

c) if the TREM, e.g. if the AC stem loop of the TREM, comprises an exogenous insert, the exogenous insert does not comprise an effector entity, e.g. an effector entity having a primary sequence, a secondary or a tertiary structure-dependent biological function;

d) if the TREM, for example, if the AC stem loop of the TREM, comprises an exogenous insert, the exogenous insert does not comprise: the epsilon domain of human hepatitis b virus; a dimerization domain of HIV; or an aptamer that binds to malachite green, dextran, or streptavidin;

e) the TREM may be loaded with amino acids;

f) the TREM is translationally competent, e.g., capable of modulating elongation of a nascent polypeptide;

g) the TREM is not a naturally occurring molecule;

h) the TREM is not a native molecule with anti-angiogenic properties, e.g., as determined by inhibition of endothelial cell proliferation;

i) the TREM is not anti-angiogenic; and

j) TREM in homologous cells does not produce naturally occurring anti-angiogenic fragments.

181. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 comprising property (f).

182. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 comprising a property selected from (a) - (f) and a property selected from (g) - (j).

183. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 comprising properties (g) and/or (d).

184. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 183, further comprising property (h) or (i).

185. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of examples 180-184 comprising a property selected from the group consisting of:

a) the composition comprises at least 1, 2, 5, 10, or 1,000 grams of TREM;

b) the compositions do not comprise full length trnas and naturally occurring anti-angiogenic fragments thereof; or

c) The composition comprises a TREM as described in any one of embodiments 67-82.

186. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 or 181 comprising a property selected from (a) - (e) and a property selected from (g) - (j).

187. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 comprising any one of properties (a) - (f).

188. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 comprising any two of characteristics (a) - (f).

189. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 comprising any three of characteristics (a) - (f).

190. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 comprising any four of characteristics (a) - (f).

191. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 comprising any five of characteristics (a) - (f).

192. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 comprising all of characteristics (a) - (f).

193. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 comprising any one of characteristics (f) - (j).

194. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 comprising any two of characteristics (f) - (j).

195. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 comprising any three of characteristics (f) - (j).

196. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 comprising any four of characteristics (f) - (j).

197. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 comprising all of characteristics (f) - (j).

198. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 180 further comprising any one, two, three or all of characteristics (g) - (j).

199. The composition or the pharmaceutical composition as defined in any one of examples 67-82 or 139-198, the method as defined in any one of examples 83-109 or 139-198, the cell as defined in any one of examples 110-132 or 139-198, the reaction mixture as defined in example 133 or 139-198, the bioreactor as defined in example 134-135 or 139-198, the master cell library as defined in example 136-137 or 139-198, or the evaluation method as defined in example 138 or 139-198, wherein the TREM recognizes the stop codon.

200. The composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of example 199, wherein the TREM mediates acceptance and incorporation of amino acids.

201. The composition or the pharmaceutical composition as defined in any one of examples 67-82 or 139-198, the method as defined in any one of examples 83-109 or 139-198, the cell as defined in any one of examples 110-132 or 139-198, the reaction mixture as defined in example 133 or 139-198, the bioreactor as defined in example 134-135 or 139-198, the master cell library as defined in example 136-137 or 139-198, or the evaluation method as defined in example 138 or 139-198, wherein the TREM does not recognize a stop codon.

202. The method of any one of examples 1-66 or 139-.

203. The method of any one of examples 1-66 or 139-198, the composition or the pharmaceutical composition of any one of examples 67-82 or 139-201, the method of any one of examples 83-109 or 139-201, the cell of any one of examples 110-132 or 139-201, the reaction mixture of example 133 or 139-201, the bioreactor of example 134-135 or 139-201, the main cell library of example 136-137 or 139-201, or the method of assessing of example 138 or 139-201, wherein the TREM is formulated as a lyophilized TREM composition.

204. The method of any one of examples 1-66 or 139-198, the composition or the pharmaceutical composition of any one of examples 67-82 or 139-201, the method of any one of examples 83-109 or 139-201, the cell of any one of examples 110-132 or 139-201, the reaction mixture of example 133 or 139-201, the bioreactor of example 134-135 or 139-201, the main cell library of example 136-137 or 139-201, or the method of assessing of example 138 or 139-201, wherein the TREM is formulated as a liquid TREM composition.

205. The method of any one of examples 1-66 or 139-198, the composition or the pharmaceutical composition of any one of examples 67-82 or 139-201, the method of any one of examples 83-109 or 139-201, the cell of any one of examples 110-132 or 139-201, the reaction mixture of example 133 or 139-201, the bioreactor of example 134-135 or 139-201, the main cell bank of example 136-137 or 139-201, or the method of assessing of example 138 or 139-201, wherein the TREM is formulated as a frozen TREM composition.

206. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:1, or a fragment thereof.

207. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:2, or a fragment thereof.

208. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 3, or a fragment thereof.

209. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 4, or a fragment thereof.

210. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 5, or a fragment thereof.

211. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 6, or a fragment thereof.

212. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 7, or a fragment thereof.

213. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:8, or a fragment thereof.

214. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:9, or a fragment thereof.

215. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:10, or a fragment thereof.

216. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:11, or a fragment thereof.

217. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 12, or a fragment thereof.

218. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:13, or a fragment thereof.

219. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 14, or a fragment thereof.

220. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:15, or a fragment thereof.

221. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:16, or a fragment thereof.

222. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:17, or a fragment thereof.

223. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:18, or a fragment thereof.

224. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:19, or a fragment thereof.

225. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:20, or a fragment thereof.

226. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:21, or a fragment thereof.

227. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:22, or a fragment thereof.

228. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 23, or a fragment thereof.

229. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:24, or a fragment thereof.

230. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:25, or a fragment thereof.

231. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:26, or a fragment thereof.

232. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:27, or a fragment thereof.

233. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:28, or a fragment thereof.

234. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:29, or a fragment thereof.

235. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:30, or a fragment thereof.

236. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:31, or a fragment thereof.

237. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:32, or a fragment thereof.

238. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:33, or a fragment thereof.

239. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:34, or a fragment thereof.

240. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:35, or a fragment thereof.

241. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:36, or a fragment thereof.

242. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:37, or a fragment thereof.

243. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:38, or a fragment thereof.

244. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:39, or a fragment thereof.

245. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:40, or a fragment thereof.

246. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:41, or a fragment thereof.

247. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:42, or a fragment thereof.

248. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:43, or a fragment thereof.

249. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:44, or a fragment thereof.

250. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:45, or a fragment thereof.

251. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:46, or a fragment thereof.

252. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:47, or a fragment thereof.

253. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:48, or a fragment thereof.

254. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:49, or a fragment thereof.

255. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:50, or a fragment thereof.

256. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:51, or a fragment thereof.

257. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:52, or a fragment thereof.

258. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:53, or a fragment thereof.

259. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:54, or a fragment thereof.

260. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:55, or a fragment thereof.

261. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:56, or a fragment thereof.

262. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:57, or a fragment thereof.

263. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:58, or a fragment thereof.

264. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:59, or a fragment thereof.

265. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:60, or a fragment thereof.

266. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:61, or a fragment thereof.

267. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:62, or a fragment thereof.

268. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:63, or a fragment thereof.

269. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:64, or a fragment thereof.

270. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:65, or a fragment thereof.

271. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:66, or a fragment thereof.

272. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:67, or a fragment thereof.

273. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:68, or a fragment thereof.

274. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:69, or a fragment thereof.

275. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:70, or a fragment thereof.

276. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:71, or a fragment thereof.

277. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:72, or a fragment thereof.

278. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 73, or a fragment thereof.

279. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:74, or a fragment thereof.

280. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:75, or a fragment thereof.

281. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:76, or a fragment thereof.

282. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:77, or a fragment thereof.

283. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:78, or a fragment thereof.

284. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:79, or a fragment thereof.

285. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:80, or a fragment thereof.

286. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:81, or a fragment thereof.

287. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:82, or a fragment thereof.

288. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:83, or a fragment thereof.

289. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:84, or a fragment thereof.

290. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:85, or a fragment thereof.

291. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:86, or a fragment thereof.

292. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:87, or a fragment thereof.

293. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:88, or a fragment thereof.

294. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 89, or a fragment thereof.

295. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:90, or a fragment thereof.

296. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:91, or a fragment thereof.

297. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:92, or a fragment thereof.

298. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:93, or a fragment thereof.

299. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:94, or a fragment thereof.

300. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:95, or a fragment thereof.

301. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:96, or a fragment thereof.

302. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:97, or a fragment thereof.

303. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:98, or a fragment thereof.

304. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 99, or a fragment thereof.

305. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:100, or a fragment thereof.

306. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:101, or a fragment thereof.

307. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:102, or a fragment thereof.

308. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 103, or a fragment thereof.

309. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:104, or a fragment thereof.

310. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:105, or a fragment thereof.

311. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:106, or a fragment thereof.

312. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:107, or a fragment thereof.

313. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:108, or a fragment thereof.

314. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:109, or a fragment thereof.

315. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:110, or a fragment thereof.

316. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 111, or a fragment thereof.

317. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:112, or a fragment thereof.

318. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 113, or a fragment thereof.

319. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:114, or a fragment thereof.

320. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:115, or a fragment thereof.

321. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:116, or a fragment thereof.

322. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:117, or a fragment thereof.

323. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:118, or a fragment thereof.

324. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:119, or a fragment thereof.

325. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:120, or a fragment thereof.

326. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 121, or a fragment thereof.

327. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 122, or a fragment thereof.

328. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 123, or a fragment thereof.

329. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:124, or a fragment thereof.

330. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:125, or a fragment thereof.

331. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:126, or a fragment thereof.

332. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:127, or a fragment thereof.

333. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:128, or a fragment thereof.

334. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:129, or a fragment thereof.

335. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:130, or a fragment thereof.

336. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:131, or a fragment thereof.

337. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:132, or a fragment thereof.

338. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:133, or a fragment thereof.

339. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:134, or a fragment thereof.

340. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:135, or a fragment thereof.

341. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:136, or a fragment thereof.

342. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:137, or a fragment thereof.

343. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:138, or a fragment thereof.

344. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:139, or a fragment thereof.

345. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:140, or a fragment thereof.

346. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:141, or a fragment thereof.

347. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:142, or a fragment thereof.

348. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 143, or a fragment thereof.

349. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:144, or a fragment thereof.

350. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:145, or a fragment thereof.

351. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:146, or a fragment thereof.

352. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:147, or a fragment thereof.

353. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 148, or a fragment thereof.

354. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:149, or a fragment thereof.

355. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:150, or a fragment thereof.

356. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:151, or a fragment thereof.

357. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:152, or a fragment thereof.

358. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:153, or a fragment thereof.

359. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:154, or a fragment thereof.

360. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:155, or a fragment thereof.

361. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:156, or a fragment thereof.

362. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:157, or a fragment thereof.

363. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:158, or a fragment thereof.

364. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:159, or a fragment thereof.

365. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:160, or a fragment thereof.

366. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:161, or a fragment thereof.

367. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:162, or a fragment thereof.

368. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 163, or a fragment thereof.

369. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:164, or a fragment thereof.

370. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:165, or a fragment thereof.

371. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:166, or a fragment thereof.

372. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:167, or a fragment thereof.

373. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:168, or a fragment thereof.

374. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 169, or a fragment thereof.

375. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:170, or a fragment thereof.

376. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 171, or a fragment thereof.

377. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:172, or a fragment thereof.

378. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 173, or a fragment thereof.

379. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:174, or a fragment thereof.

380. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:175, or a fragment thereof.

381. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:176, or a fragment thereof.

382. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:177, or a fragment thereof.

383. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:178, or a fragment thereof.

384. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:179, or a fragment thereof.

385. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:180, or a fragment thereof.

386. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:181, or a fragment thereof.

387. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 182, or a fragment thereof.

388. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:183, or a fragment thereof.

389. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:184, or a fragment thereof.

390. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:185, or a fragment thereof.

391. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:186, or a fragment thereof.

392. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 187, or a fragment thereof.

393. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:188, or a fragment thereof.

394. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:189, or a fragment thereof.

395. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:190, or a fragment thereof.

396. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:191, or a fragment thereof.

397. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:192, or a fragment thereof.

398. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:193, or a fragment thereof.

399. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:194, or a fragment thereof.

400. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:195, or a fragment thereof.

401. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:196, or a fragment thereof.

402. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:197, or a fragment thereof.

403. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:198, or a fragment thereof.

404. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:199, or a fragment thereof.

405. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:200, or a fragment thereof.

406. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:201, or a fragment thereof.

407. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:202, or a fragment thereof.

408. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 203, or a fragment thereof.

409. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:204, or a fragment thereof.

410. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 205, or a fragment thereof.

411. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:206, or a fragment thereof.

412. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:207, or a fragment thereof.

413. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:208, or a fragment thereof.

414. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 209, or a fragment thereof.

415. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:210, or a fragment thereof.

416. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:211, or a fragment thereof.

417. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:212, or a fragment thereof.

418. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:213, or a fragment thereof.

419. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:214, or a fragment thereof.

420. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:215, or a fragment thereof.

421. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:216, or a fragment thereof.

422. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 217, or a fragment thereof.

423. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 218, or a fragment thereof.

424. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:219, or a fragment thereof.

425. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:220, or a fragment thereof.

426. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 221, or a fragment thereof.

427. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:222, or a fragment thereof.

428. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:223, or a fragment thereof.

429. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:224, or a fragment thereof.

430. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:225, or a fragment thereof.

431. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:226, or a fragment thereof.

432. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:227, or a fragment thereof.

433. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:228, or a fragment thereof.

434. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 229, or a fragment thereof.

435. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:230, or a fragment thereof.

436. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:231, or a fragment thereof.

437. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:232, or a fragment thereof.

438. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 233, or a fragment thereof.

439. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:234, or a fragment thereof.

440. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:235, or a fragment thereof.

441. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:236, or a fragment thereof.

442. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:237, or a fragment thereof.

443. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:238, or a fragment thereof.

444. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:239, or a fragment thereof.

445. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:240, or a fragment thereof.

446. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:241, or a fragment thereof.

447. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:242, or a fragment thereof.

448. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:243, or a fragment thereof.

449. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 244, or a fragment thereof.

450. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:245, or a fragment thereof.

451. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 246, or a fragment thereof.

452. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 247, or a fragment thereof.

453. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:248, or a fragment thereof.

454. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:249, or a fragment thereof.

455. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:250, or a fragment thereof.

456. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:251, or a fragment thereof.

457. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:252, or a fragment thereof.

458. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:253, or a fragment thereof.

459. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:254, or a fragment thereof.

460. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:255, or a fragment thereof.

461. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:256, or a fragment thereof.

462. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:257, or a fragment thereof.

463. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:258, or a fragment thereof.

464. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:259, or a fragment thereof.

465. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:260, or a fragment thereof.

466. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:261, or a fragment thereof.

467. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 262, or a fragment thereof.

468. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:263, or a fragment thereof.

469. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:264, or a fragment thereof.

470. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:265, or a fragment thereof.

471. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 266, or a fragment thereof.

472. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 267, or a fragment thereof.

473. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:268, or a fragment thereof.

474. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:269, or a fragment thereof.

475. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:270, or a fragment thereof.

476. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 271, or a fragment thereof.

477. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:272, or a fragment thereof.

478. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:273, or a fragment thereof.

479. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:274, or a fragment thereof.

480. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:275, or a fragment thereof.

481. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:276, or a fragment thereof.

482. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 277, or a fragment thereof.

483. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 278, or a fragment thereof.

484. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:279, or a fragment thereof.

485. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:280, or a fragment thereof.

486. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 281, or a fragment thereof.

487. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 282, or a fragment thereof.

488. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:283, or a fragment thereof.

489. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:284, or a fragment thereof.

490. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:285, or a fragment thereof.

491. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 286, or a fragment thereof.

492. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:287, or a fragment thereof.

493. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:288, or a fragment thereof.

494. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:289, or a fragment thereof.

495. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:290, or a fragment thereof.

496. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:291, or a fragment thereof.

497. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:292, or a fragment thereof.

498. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:293, or a fragment thereof.

499. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:294, or a fragment thereof.

500. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:295, or a fragment thereof.

501. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:296, or a fragment thereof.

502. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:297, or a fragment thereof.

503. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:298, or a fragment thereof.

504. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:299, or a fragment thereof.

505. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:300, or a fragment thereof.

506. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:301, or a fragment thereof.

507. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:302, or a fragment thereof.

508. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 303, or a fragment thereof.

509. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:304, or a fragment thereof.

510. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:305, or a fragment thereof.

511. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:306, or a fragment thereof.

512. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:307, or a fragment thereof.

513. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:308, or a fragment thereof.

514. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:309, or a fragment thereof.

515. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:310, or a fragment thereof.

516. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:311, or a fragment thereof.

517. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:312, or a fragment thereof.

518. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:313, or a fragment thereof.

519. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:314, or a fragment thereof.

520. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 315, or a fragment thereof.

521. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:316, or a fragment thereof.

522. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:317, or a fragment thereof.

523. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:318, or a fragment thereof.

524. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:319, or a fragment thereof.

525. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:320, or a fragment thereof.

526. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 321, or a fragment thereof.

527. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:322, or a fragment thereof.

528. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:323, or a fragment thereof.

529. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:324, or a fragment thereof.

530. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:325, or a fragment thereof.

531. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:326, or a fragment thereof.

532. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:327, or a fragment thereof.

533. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:328, or a fragment thereof.

534. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:329, or a fragment thereof.

535. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:330, or a fragment thereof.

536. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:331, or a fragment thereof.

537. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:332, or a fragment thereof.

538. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:333, or a fragment thereof.

539. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:334, or a fragment thereof.

540. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:335, or a fragment thereof.

541. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:336, or a fragment thereof.

542. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 337, or a fragment thereof.

543. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:338, or a fragment thereof.

544. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:339, or a fragment thereof.

545. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:340, or a fragment thereof.

546. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:341, or a fragment thereof.

547. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:342, or a fragment thereof.

548. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:343, or a fragment thereof.

549. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 344, or a fragment thereof.

550. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:345, or a fragment thereof.

551. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 346, or a fragment thereof.

552. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:347, or a fragment thereof.

553. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:348, or a fragment thereof.

554. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:349, or a fragment thereof.

555. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:350, or a fragment thereof.

556. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:351, or a fragment thereof.

557. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:352, or a fragment thereof.

558. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:353, or a fragment thereof.

559. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:354, or a fragment thereof.

560. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:355, or a fragment thereof.

561. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:356, or a fragment thereof.

562. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:357, or a fragment thereof.

563. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:358, or a fragment thereof.

564. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:359, or a fragment thereof.

565. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:360, or a fragment thereof.

566. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:361, or a fragment thereof.

567. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:362, or a fragment thereof.

568. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:363, or a fragment thereof.

569. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:364, or a fragment thereof.

570. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 365, or a fragment thereof.

571. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 366, or a fragment thereof.

572. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No.: 367, or a fragment thereof.

573. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 368, or a fragment thereof.

574. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 369, or a fragment thereof.

575. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 370, or a fragment thereof.

576. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:371, or a fragment thereof.

577. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:372, or a fragment thereof.

578. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:373, or a fragment thereof.

579. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:374, or a fragment thereof.

580. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:375, or a fragment thereof.

581. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 376, or a fragment thereof.

582. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:377, or a fragment thereof.

583. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:378, or a fragment thereof.

584. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:379, or a fragment thereof.

585. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:380, or a fragment thereof.

586. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 381, or a fragment thereof.

587. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:382, or a fragment thereof.

588. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 383, or a fragment thereof.

589. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:384, or a fragment thereof.

590. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:385, or a fragment thereof.

591. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:386, or a fragment thereof.

592. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:387, or a fragment thereof.

593. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:388, or a fragment thereof.

594. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:389, or a fragment thereof.

595. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:390, or a fragment thereof.

596. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:391, or a fragment thereof.

597. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:392, or a fragment thereof.

598. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:393, or a fragment thereof.

599. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:394, or a fragment thereof.

600. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 395, or a fragment thereof.

601. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:396, or a fragment thereof.

602. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:397, or a fragment thereof.

603. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 398, or a fragment thereof.

604. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:399, or a fragment thereof.

605. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:400, or a fragment thereof.

606. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:401, or a fragment thereof.

607. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:402, or a fragment thereof.

608. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:403, or a fragment thereof.

609. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:404, or a fragment thereof.

610. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:405, or a fragment thereof.

611. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:406, or a fragment thereof.

612. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:407, or a fragment thereof.

613. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:408, or a fragment thereof.

614. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:409, or a fragment thereof.

615. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:410, or a fragment thereof.

616. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:411, or a fragment thereof.

617. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 412, or a fragment thereof.

618. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:413, or a fragment thereof.

619. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 414, or a fragment thereof.

620. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 415, or a fragment thereof.

621. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:416, or a fragment thereof.

622. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:417, or a fragment thereof.

623. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:418, or a fragment thereof.

624. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 419, or a fragment thereof.

625. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:420, or a fragment thereof.

626. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:421, or a fragment thereof.

627. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:422, or a fragment thereof.

628. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:423, or a fragment thereof.

629. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:424, or a fragment thereof.

630. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:425, or a fragment thereof.

631. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:426, or a fragment thereof.

632. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 427, or a fragment thereof.

633. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:428, or a fragment thereof.

634. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 429, or a fragment thereof.

635. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:430, or a fragment thereof.

636. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:431, or a fragment thereof.

637. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 432, or a fragment thereof.

638. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:433, or a fragment thereof.

639. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:434, or a fragment thereof.

640. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:435, or a fragment thereof.

641. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:436, or a fragment thereof.

642. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 437, or a fragment thereof.

643. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:438, or a fragment thereof.

644. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:439, or a fragment thereof.

645. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:440, or a fragment thereof.

646. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:441, or a fragment thereof.

647. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:442, or a fragment thereof.

648. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:443, or a fragment thereof.

649. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:444, or a fragment thereof.

650. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:445, or a fragment thereof.

651. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:446, or a fragment thereof.

652. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:447, or a fragment thereof.

653. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 448, or a fragment thereof.

654. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:449, or a fragment thereof.

655. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:450, or a fragment thereof.

656. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:451, or a fragment thereof.

657. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:562, or a fragment thereof.

658. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:563, or a fragment thereof.

659. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:564, or a fragment thereof.

660. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:565, or a fragment thereof.

661. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:566, or a fragment thereof.

662. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:567, or a fragment thereof.

663. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:568, or a fragment thereof.

664. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:569, or a fragment thereof.

665. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:570, or a fragment thereof.

666. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:571, or a fragment thereof.

667. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:572, or a fragment thereof.

668. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:573, or a fragment thereof.

669. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 574, or a fragment thereof.

670. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:575, or a fragment thereof.

671. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:576, or a fragment thereof.

672. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 577, or a fragment thereof.

673. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:578, or a fragment thereof.

674. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 579, or a fragment thereof.

675. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:580, or a fragment thereof.

676. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:581, or a fragment thereof.

677. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:582, or a fragment thereof.

678. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:583, or a fragment thereof.

679. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:584, or a fragment thereof.

680. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:585, or a fragment thereof.

681. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:586, or a fragment thereof.

682. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:587, or a fragment thereof.

683. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:588, or a fragment thereof.

684. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:589, or a fragment thereof.

685. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:590, or a fragment thereof.

686. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:591, or a fragment thereof.

687. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:592, or a fragment thereof.

688. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:593, or a fragment thereof.

689. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:594, or a fragment thereof.

690. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 595, or a fragment thereof.

691. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:596, or a fragment thereof.

692. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 597, or a fragment thereof.

693. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 598, or a fragment thereof.

694. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 599, or a fragment thereof.

695. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:600, or a fragment thereof.

696. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:601, or a fragment thereof.

697. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:602, or a fragment thereof.

698. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 603, or a fragment thereof.

699. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:604, or a fragment thereof.

700. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:605, or a fragment thereof.

701. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:606, or a fragment thereof.

702. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 607, or a fragment thereof.

703. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 608, or a fragment thereof.

704. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 609, or a fragment thereof.

705. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:610, or a fragment thereof.

706. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:611, or a fragment thereof.

707. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:612, or a fragment thereof.

708. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID No. 613, or a fragment thereof.

709. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:614, or a fragment thereof.

710. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:615, or a fragment thereof.

711. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:616, or a fragment thereof.

712. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell bank of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:617, or a fragment thereof.

713. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:618, or a fragment thereof.

714. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 619, or a fragment thereof.

715. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO:620, or a fragment thereof.

716. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 1-205, wherein the TREM comprises an RNA sequence encoded by the DNA sequence of SEQ ID NO 621, or a fragment thereof.

717. The method, composition or pharmaceutical composition, cell, reaction mixture, bioreactor or master cell library of any one of embodiments 206-716, wherein the fragments comprise one or more, but not all of: linker 1 region, AStD stem region; a linker 2 region; stem-loop regions, e.g., the D-arm region; a linker 3 region; stem-loop regions, e.g., AC arm regions; a variable region; stem-loop regions, e.g., T-arm regions; and linker 4 regions, e.g., such as those described herein.

718. A method of making a purified tRNA effector molecule (TREM) pharmaceutical composition, the method comprising:

providing an insect host cell comprising an exogenous nucleic acid, e.g., DNA or RNA, encoding a TREM;

maintaining the insect host cell under conditions sufficient to express a TREM;

purifying TREM from the insect host cell, e.g., according to the methods described herein; and

formulating the purified TREM into a pharmaceutical composition, e.g. by combining said TREM with a pharmaceutical excipient,

Thereby preparing the TREM pharmaceutical composition.

719. The method of embodiment 718, wherein the insect host cell is selected from the group consisting of: insect cells or cell lines, for example, Sf9 cells or cell lines.

720. A method of making a purified tRNA effector molecule (TREM) pharmaceutical composition, the method comprising:

providing a yeast host cell comprising an exogenous nucleic acid, e.g., DNA or RNA, encoding a TREM;

maintaining the yeast host cell under conditions sufficient to express a TREM;

purifying TREM from the yeast host cell, e.g., according to the methods described herein; and

formulating the purified TREM into a pharmaceutical composition, e.g. by combining said TREM with a pharmaceutical excipient,

thereby preparing the TREM pharmaceutical composition.

721. The method of embodiment 720, wherein the yeast host cell is selected from the group consisting of: yeast cells or cell lines, for example Saccharomyces cerevisiae or Schizosaccharomyces pombe cells or cell lines.

722. The method of any one of embodiments 718-721, wherein the purifying step comprises one, two or all of the following steps, e.g., in the order recited:

(i) separating the nucleic acids from the proteins to provide an RNA preparation;

(ii) Separating RNA less than a threshold number of nucleotides, e.g., less than 500nt, less than 400nt, less than 300nt, less than 250nt, less than 200nt, less than 150nt, from larger RNA species in the RNA preparation to produce a small RNA preparation; and/or

(iii) TREM is separated from other RNA species in the small RNA preparation by affinity-based separation, e.g., sequence affinity.

Other features, objects, and advantages of the invention will be apparent from the description and from the claims.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

Drawings

FIGS. 1A-1C are graphs showing increased cell growth in three cell lines following transfection with a TREM corresponding to the initiator methionine (iMet). Figure 1A is a graph showing increased cell confluence% (measure of cell growth) of U20S cells transfected with either met-CAT-TREM labeled with Cy3 or with non-targeted control labeled with Cy 3. Figure 1B is a graph showing increased cell confluence% (measure of cell growth) of H1299 cells transfected with either a Cy 3-labeled iMet-CAT-TREM or a Cy 3-labeled non-targeted control. FIG. 1C is a graph showing increased cell confluence% (measure of cell growth) of Hela cells transfected with Cy 3-labeled iMet-CAT-TREM or with Cy 3-labeled non-targeted control.

Figure 2 is a graph depicting increased NanoLuc reporter expression following addition of an iMET-TREM to a translation reaction with cell-free lysate. As a control, translation reaction was performed with buffer.

Detailed Description

The disclosure features tRNA-based effector molecules (TREMs) and methods related thereto. tRNA-based effector molecules (TREMs) as disclosed herein are complex molecules that can mediate a variety of cellular processes. The pharmaceutical TREM composition can be administered to a cell, tissue, or subject to modulate these functions.

Definition of

The term "homologous adaptor function TREM" as used herein refers to TREM which mediates the initiation or extension of an AA (homologous AA) naturally associated with the anticodon of TREM.

The term "reduced expression" as used herein refers to a reduction in expression as compared to a reference, e.g., where an altered control region or addition of an agent results in reduced expression of a product in a subject, relative to an otherwise similar cell without the alteration or addition.

The term "exogenous nucleic acid" as used herein refers to a nucleic acid sequence that is not present in a reference cell, e.g., a cell into which the exogenous nucleic acid is introduced, or that differs from the closest sequence in the reference cell by at least one nucleotide. In embodiments, the exogenous nucleic acid comprises a nucleic acid encoding a TREM.

The term "exogenous TREM" as used herein refers to a TREM having the following:

(a) at least one nucleotide or one post-transcriptional modification that differs from the closest sequence tRNA in a reference cell, e.g., a cell into which an exogenous nucleic acid is introduced;

(b) cells other than the cells into which they have been introduced for transcription;

(d) Has an expression profile, e.g., level or profile, which is not wild-type, e.g., its expression level is higher than wild-type. In embodiments, expression profiling can be mediated by introducing changes into nucleic acids that modulate expression or by adding agents that modulate expression of RNA molecules. In embodiments, the exogenous TREM comprises 1, 2, 3, or 4 of the properties (a) - (d).

The term "GMP-grade composition" as used herein refers to a composition that meets current good manufacturing practice (cGMP) guidelines or other similar requirements. In an embodiment, the GMP-grade composition may be used as a pharmaceutical product.

As used herein, the terms "increase" and "decrease" refer to modulating a function, expression, or activity, respectively, that produces a particular indicator by a greater or lesser amount relative to a reference. For example, following administration of a TREM as described herein to a cell, tissue, or subject, the amount of a marker for an indicator as described herein (e.g., protein translation, mRNA stability, protein folding) can be increased or decreased by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%, 2X, 3X, 5X, 10X, or more, relative to the amount of the marker prior to administration, or relative to the effect of a negative control agent. The post-administration indicator is measured at a time when the administration has achieved the effect, e.g., at least 12 hours, 24 hours, one week, one month, 3 months, or 6 months after the start of treatment.

The term "increased expression" as used herein refers to an increase in expression as compared to a reference, e.g., where an altered control region or addition of an agent results in increased expression of a product in a subject, relative to an otherwise similar cell without the alteration or addition.

The term "non-homologous adaptor function TREM" as used herein refers to TREM that mediates the initiation or extension of an AA other than the AA naturally associated with the anticodon of TREM (a non-homologous AA). In embodiments, the non-homologous adaptor-functional TREM is also referred to as unloaded TREM (mtrem).

The term "non-naturally occurring sequence" as used herein refers to a sequence in which adenine is replaced with a residue other than an adenine analog, cytosine is replaced with a residue other than a cytosine analog, guanine is replaced with a residue other than a guanine analog, and uracil is replaced with a residue other than a uracil analog. Analogs refer to any possible derivative of ribonucleotide A, G, C or U. In embodiments, the sequence having a derivative of either ribonucleotide A, G, C or U is a non-naturally occurring sequence.

As used herein, "oncogene" refers to a gene that modulates one or more cellular processes, including: cell fate determination, cell survival and genome maintenance. In embodiments, the oncogene provides a selective growth advantage to the cell in which it is present, e.g., a disorder, e.g., a genetic disorder (e.g., mutation or amplification) or an epigenetic disorder. Exemplary oncogenes include Myc (e.g., c-Myc, N-Myc or L-Myc), c-Jun, Wnt or RAS.

The term "pharmaceutical TREM composition" as used herein refers to a TREM composition suitable for pharmaceutical use. Typically, a pharmaceutical TREM composition comprises a pharmaceutical excipient. In the examples, TREM will be the only active ingredient in the pharmaceutical TREM composition. In embodiments, the pharmaceutical TREM composition is free, substantially free, or has less than a pharmaceutically acceptable amount of host cell proteins, DNA, e.g., host cell DNA, endotoxins, and bacteria.

The term "post-transcriptional processing" as used herein with respect to a subject molecule, e.g., a TREM, RNA, or tRNA, refers to covalent modification of the subject molecule. In embodiments, the covalent modification occurs post-transcriptionally. In embodiments, the covalent modification occurs co-transcriptionally. In embodiments, the modification is performed in vivo, e.g., in a cell used to produce TREM. In embodiments, the modification is in vitro, e.g., it is performed on TREM isolated or obtained from a cell producing TREM. In embodiments, the post-transcriptional modification is selected from the post-transcriptional modifications listed in table 2.

The term "recombinant TREM" as used herein refers to a TREM that is expressed in a cell that has been modified by human intervention with a modification that mediates TREM production, e.g., the cell comprises an exogenous sequence that encodes the TREM, or a modification that mediates expression, e.g., transcriptional expression or post-transcriptional modification of the TREM. The recombinant TREM can have the same or different sequence, post-transcriptional modification group, or tertiary structure as the reference tRNA, e.g., a native tRNA.

The term "synthesized TREM" as used herein refers to TREM synthesized (e.g., by cell-free solid phase synthesis) in a cell other than a cell having an endogenous nucleic acid encoding TREM. The synthetic TREM can have the same or different sequence, post-transcriptional modification group, or tertiary structure as the native tRNA.

The term "TREM expressed in a heterologous cell" as used herein refers to TREM prepared under non-native conditions. For example, TREM, i) is produced in a cell that is different from a naturally occurring cell, e.g., genetically, metabolically (e.g., has a different gene expression profile or has different levels of cellular components, e.g., absorbed nutrients), or epigenetically different; ii) produced in a cell cultured under conditions different from native conditions (native conditions being conditions under which the cell produces tRNA in nature), e.g., nutrient, pH, temperature, cell density, or pressure conditions; or iii) is manufactured at a different level, rate or concentration in the cell than the reference, or is positioned in a different compartment or location than the reference, e.g., is manufactured at a different level, rate or concentration than occurs under native conditions, or is positioned in a different compartment or location than occurs under native conditions. The TREM expressed in the heterologous cell can have the same or different sequence, post-transcriptional modification group, or tertiary structure as the native tRNA.

The term "tRNA" as used herein refers to a naturally occurring transfer ribonucleic acid in its natural state.

The term "tRNA-based effector molecule" or "TREM" as used herein refers to an RNA molecule comprising a structure or characteristic from (a) - (v) below, and which is a recombinant TREM, a synthetic TREM, or a TREM expressed from a heterologous cell. A TREM can have a plurality (e.g., 2, 3, 4, 5, 6, 7, 8, 9) of structures and functions in (a) - (v).

In embodiments, TREM is non-native as assessed by its structure or manner of preparation.

In an embodiment, TREM includes one or more of the following structures or characteristics:

(a') an optional linker region, e.g., linker 1 region, of the consensus sequence provided in the "consensus sequence" section;

(a) an amino acid attachment domain that binds an amino acid, e.g., an acceptor stem domain (AStD), wherein the AStD comprises sufficient RNA sequence to mediate, e.g., accept, transfer of an amino acid, e.g., a homologous or nonhomologous amino acid thereof, and an Amino Acid (AA) in the initiation or extension of a polypeptide chain, e.g., when present in an otherwise wild-type tRNA. Typically, the AStD contains a 3' -terminal adenosine (CCA) for acceptance of the stem load, which is part of the recognition by the synthetase. In embodiments, the AStD is at least 75%, 80%, 85%, 90%, 95%, or 100% identical to a naturally occurring AStD (e.g., an AStD encoded by a nucleic acid in table 1). In embodiments, a TREM may comprise a fragment or analog of an AStD (e.g., an AStD encoded by a nucleic acid in table 1) that has, in embodiments, AStD activity and, in other embodiments, does not have AStD activity. (the ordinarily skilled artisan can determine the relevant corresponding sequences for any of the domains, stems, loops, or other sequence features mentioned herein from the sequences encoded by the nucleic acids in Table 1. for example, the ordinarily skilled artisan can determine the sequences corresponding to AStD from the tRNA sequences encoded by the nucleic acids in Table 1.)

In embodiments, the AStD belongs to the corresponding sequence of the consensus sequence provided in the "consensus sequence" section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions;

in embodiments, the AStD comprises formula I_ZZZResidue R of₁-R₂-R₃-R₄-R₅-R₆-R₇And a residue R₆₅-R₆₆-R₆₇-R₆₈-R₆₉-R₇₀-R₇₁Wherein ZZZ represents any one of twenty amino acids;

in embodiments, the AStD comprises formula II_ZZZResidue R of₁-R₂-R₃-R₄-R₅-R₆-R₇And a residue R₆₅-R₆₆-R₆₇-R₆₈-R₆₉-R₇₀-R₇₁Wherein ZZZ represents any one of twenty amino acids;

in embodiments, the AStD comprises formula III_ZZZResidue R of₁-R₂-R₃-R₄-R₅-R₆-R₇And a residue R₆₅-R₆₆-R₆₇-R₆₈-R₆₉-R₇₀-R₇₁Wherein ZZZ represents any one of twenty amino acids;

(a' -1) residue R comprising the consensus sequence provided in the "consensus sequence" section₈-R₉The linker of (a), e.g., linker 2 region;

(b) a Dihydrouridine Hairpin Domain (DHD), wherein the DHD comprises sufficient RNA sequence to mediate, e.g., recognize, e.g., serve as a recognition site for, an aminoacyl-tRNA synthetase when present in an otherwise wild-type tRNA, for amino acid charging of TREM. In embodiments, DHD mediates stabilization of the TREM tertiary structure. In embodiments, the DHD is at least 75%, 80%, 85%, 90%, 95%, or 100% identical to a naturally occurring DHD (e.g., a DHD encoded by a nucleic acid in table 1). In embodiments, a TREM may comprise a fragment or analog of a DHD (e.g., a DHD encoded by a nucleic acid in table 1), which fragment has DHD activity in embodiments and does not have DHD activity in other embodiments.

In embodiments, the DHD belongs to the corresponding sequence of the consensus sequence provided in the "consensus sequence" section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions;

in embodiments, the DHD comprises formula I_ZZZResidue R of₁₀-R₁₁-R₁₂-R₁₃-R₁₄R₁₅-R₁₆-R₁₇-R₁₈-R₁₉-R₂₀-R₂₁-R₂₂-R₂₃-R₂₄-R₂₅-R₂₆-R₂₇-R₂₈Wherein ZZZ represents any one of twenty amino acids;

in embodiments, the DHD comprises formula II_ZZZResidue R of₁₀-R₁₁-R₁₂-R₁₃-R₁₄R₁₅-R₁₆-R₁₇-R₁₈-R₁₉-R₂₀-R₂₁-R₂₂-R₂₃-R₂₄-R₂₅-R₂₆-R₂₇-R₂₈Wherein ZZZ represents any one of twenty amino acids;

in embodiments, the DHD comprises formula III_ZZZResidue R of₁₀-R₁₁-R₁₂-R₁₃-R₁₄R₁₅-R₁₆-R₁₇-R₁₈-R₁₉-R₂₀-R₂₁-R₂₂-R₂₃-R₂₄-R₂₅-R₂₆-R₂₇-R₂₈Wherein ZZZ represents any one of twenty amino acids;

(b' -1) residue R comprising the consensus sequence provided in the "consensus sequence" section₂₉The linker of (a), e.g., linker 3 region;

(c) an anticodon that binds to a corresponding codon in the mRNA, e.g., an anticodon hairpin domain (ACHD), wherein the ACHD comprises sufficient sequence, e.g., an anticodon triplet, to mediate pairing (with or without wobble) with the codon, e.g., when present in an otherwise wild-type tRNA; in embodiments, the ACHD is at least 75%, 80%, 85%, 90%, 95%, or 100% identical to a naturally occurring ACHD (e.g., an ACHD encoded by a nucleic acid in table 1). In embodiments, a TREM can comprise a fragment or analog of ACHD (e.g., ACHD encoded by a nucleic acid in table 1) that has in embodiments and in other embodiments does not have ACHD activity.

In embodiments, ACHD belongs to the corresponding sequence of the consensus sequences provided in the "consensus sequences" section, or differs from the consensus sequences by no more than 1, 2, 5, or 10 positions;

in an embodiment, ACHD comprises formula I_ZZZresidue-R₃₀-R₃₁-R₃₂-R₃₃-R₃₄-R₃₅-R₃₆-R₃₇-R₃₈-R₃₉-R₄₀-R₄₁-R₄₂-R₄₃-R₄₄-R₄₅-R₄₆Wherein ZZZ represents any one of twenty amino acids;

in an embodiment, ACHD comprises formula II_ZZZResidue of (A) to R₃₀-R₃₁-R₃₂-R₃₃-R₃₄-R₃₅-R₃₆-R₃₇-R₃₈-R₃₉-R₄₀-R₄₁-R₄₂-R₄₃-R₄₄-R₄₅-R₄₆Wherein ZZZ represents any one of twenty amino acids;

in an embodiment, ACHD comprises formula III_ZZZResidue of (A) to R₃₀-R₃₁-R₃₂-R₃₃-R₃₄-R₃₅-R₃₆-R₃₇-R₃₈-R₃₉-R₄₀-R₄₁-R₄₂-R₄₃-R₄₄-R₄₅-R₄₆Wherein ZZZ represents any one of twenty amino acids;

(d) a Variable Loop Domain (VLD), wherein the VLD comprises sufficient RNA sequence to mediate, e.g., recognize, e.g., serve as a recognition site for, an aminoacyl-tRNA synthetase when present in an otherwise wild-type tRNA, for amino acid loading of TREM. In the examples, VLD mediates stabilization of the TREM tertiary structure. In embodiments, VLD modulates, e.g., increases, the specificity of TREM, e.g., for its cognate amino acid, e.g., VLD modulates TREM's cognate adaptor function. In embodiments, the VLD is at least 75%, 80%, 85%, 90%, 95%, or 100% identical to a naturally occurring VLD (e.g., a VLD encoded by a nucleic acid in table 1). In embodiments, a TREM can comprise a fragment of VLD (e.g., a VLD encoded by a nucleic acid in table 1) or an analog, the fragment having VLD activity in embodiments and not VLD activity in other embodiments.

In embodiments, the VLD belongs to the corresponding sequence of the consensus sequence provided in the "consensus sequence" section.

In the examples, the VLD comprises residues of the consensus sequence provided in the "consensus" portion- [ R ]₄₇]_xWhere x is 1-271 (e.g., x is 1-250, x is 1-225, x is 1-200, x is 1-175, x is 1-150, x is 1-125, x is 1-100, x is 1-75, x is 1-50, x is 1-40, x is 1-30, x is 1-29, x is 1-28, x is 1-27, x is 1-26, x is 1-25, x is 1-24, x is 1-23, x is 1-22, x is 1-21, x is 1-20, x is 1-19, x is 1-18, x is 1-17, x is 1-16, x is 1-15, x is 1-14, x is 1-10, x is 1-13, x is 1-17, x is 1-16, x is 1-15, x is 1-10, x is 1-13, x is 1-10, x is 1-10, x-1-25, x is 1-25, x is 1-15, x-1-15, x-1, x-1, x-1, x, x-10-271, x-20-271, x-30-271, x-40-271, x-50-271, x-60-271, x-70-271, x-80-271, x-100-271, x-125-271, x-150-271, x-175-271, x-200-271, x-225-271, x-1, x-2, x-3, x-4, x-5, x-6, x-7, x-8, x-9, x-10, x-11, x-12, x-13, x-14, x-15, x-16, x-18, x-28, x-20, x-15, x-15, x-15, x-15, x-15, x-15, x-15, x-15, x, x-110, x-125, x-150, x-175, x-200, x-225, x-250, or x-271);

(e) A Thymine Hairpin Domain (THD), wherein the THD comprises sufficient RNA sequence to mediate the recognition of a ribosome, e.g., serve as a recognition site for a ribosome, for example, when present in an otherwise wild-type tRNA, forming a TREM-ribosome complex during translation. In embodiments, the THD is at least 75%, 80%, 85%, 90%, 95%, or 100% identical to a naturally occurring THD (e.g., a THD encoded by a nucleic acid in table 1). In embodiments, a TREM may comprise a fragment or analog of THD (e.g., THD encoded by a nucleic acid in table 1), which fragment has THD activity in embodiments and does not have THD activity in other embodiments.

In embodiments, THD belongs to the corresponding sequence of the consensus sequence provided in the "consensus sequence" section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions;

in embodiments, the THD comprises formula I_ZZZresidue-R₄₈-R₄₉-R₅₀-R₅₁-R₅₂-R₅₃-R₅₄-R₅₅-R₅₆-R₅₇-R₅₈-R₅₉-R₆₀-R₆₁-R₆₂-R₆₃-R₆₄Wherein ZZZ represents any one of twenty amino acids;

in embodiments, the THD comprises formula II_ZZZResidue of (A) to R₄₈-R₄₉-R₅₀-R₅₁-R₅₂-R₅₃-R₅₄-R₅₅-R₅₆-R₅₇-R₅₈-R₅₉-R₆₀-R₆₁-R₆₂-R₆₃-R₆₄Wherein ZZZ represents any one of twenty amino acids;

in embodiments, the THD comprises formula III_ZZZResidue of (A) to R₄₈-R₄₉-R₅₀-R₅₁-R₅₂-R₅₃-R₅₄-R₅₅-R₅₆-R₅₇-R₅₈-R₅₉-R₆₀-R₆₁-R₆₂-R₆₃-R₆₄Wherein ZZZ represents any one of twenty amino acids;

(e' 1) residue R comprising the consensus sequence provided in the "consensus sequence" part₇₂The linker of (a), e.g., linker 4 region;

(f) under physiological conditions, it comprises a stem structure and one or more loop structures, for example 1, 2 or 3 loops. The loop may comprise a domain as described herein, e.g., a domain selected from (a) - (e). A loop may comprise one or more domains. In embodiments, the stem or loop structure is at least 75%, 80%, 85%, 90%, 95%, or 100% identical to a naturally occurring stem or loop structure (e.g., a stem or loop structure encoded by a nucleic acid in table 1). In embodiments, a TREM may comprise a fragment or analog of a stem or loop structure (e.g., a stem or loop structure encoded by a nucleic acid in table 1), which in embodiments has activity of a stem or loop structure and in other embodiments does not have activity of a stem or loop structure;

(g) tertiary structures, e.g., L-shaped tertiary structures;

(h) adaptor functions, i.e., TREM mediates acceptance of an amino acid (e.g., its cognate amino acid) and transfer of AA in the initiation or extension of a polypeptide chain;

(i) homologous adaptor function, wherein TREM mediates acceptance and incorporation of an amino acid naturally associated with the anticodon of TREM (e.g., a homologous amino acid) to initiate or extend a polypeptide chain;

(j) A non-homologous adaptor function, wherein the TREM mediates acceptance and incorporation of an amino acid other than the amino acid naturally associated with the anticodon of the TREM (e.g., a non-homologous amino acid) during initiation or elongation of a polypeptide chain;

(k) regulatory functions, e.g., epigenetic functions (e.g., gene silencing functions or signaling pathway regulatory functions), cell fate regulatory functions, mRNA stability regulatory functions, protein transduction regulatory functions, or protein compartmentalization functions;

(l) A structure that allows ribosome binding;

(m) a post-transcriptional modification, e.g., it comprises one or more modifications from table 2, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 of the modifications listed in table 2;

(n) the ability to suppress a functional characteristic of the tRNA, e.g., any of characteristics (h) - (k) possessed by the tRNA;

(o) ability to modulate cell fate;

(p) ability to modulate ribosome occupancy;

(q) ability to regulate protein translation;

(r) ability to modulate mRNA stability;

(s) the ability to modulate the folding and structure of proteins;

(t) the ability to regulate protein transduction or compartmentalization;

(u) the ability to modulate protein stability; or

(v) The ability to modulate a signaling pathway, e.g., a cellular signaling pathway.

In embodiments, the TREM comprises a full length tRNA molecule or fragment thereof.

In an embodiment, TREM includes the following characteristics: (a) - (e) of (d).

In an embodiment, TREM includes the following characteristics: (a) and (c).

In an embodiment, TREM includes the following characteristics: (a) (c) and (h).

In an embodiment, TREM includes the following characteristics: (a) (c), (h) and (b).

In an embodiment, TREM includes the following characteristics: (a) (c), (h) and (e).

In an embodiment, TREM includes the following characteristics: (a) (c), (h), (b) and (e).

In an embodiment, TREM includes the following characteristics: (a) (c), (h), (b), (e) and (g).

In an embodiment, TREM includes the following characteristics: (a) (c), (h) and (m).

In an embodiment, TREM includes the following characteristics: (a) (c), (h), (m) and (g).

In an embodiment, TREM includes the following characteristics: (a) (c), (h), (m) and (b).

In an embodiment, TREM includes the following characteristics: (a) (c), (h), (m) and (e).

In an embodiment, TREM includes the following characteristics: (a) (c), (h), (m), (g), (b) and (e).

In an embodiment, TREM includes the following characteristics: (a) (c), (h), (m), (g), (b), (e) and (q).

In an embodiment, TREM comprises:

(i) an amino acid attachment domain that binds an amino acid (e.g., an AStD as described in (a) herein); and

(ii) an anti-codon that binds to a corresponding codon in the mRNA (e.g., ACHD, as described in (c) herein).

In embodiments, the TREM comprises a flexible RNA linker providing covalent attachment of (i) to (ii).

In the examples, TREM mediates protein translation.

In embodiments, the TREM comprises a linker, e.g., an RNA linker, e.g., a flexible RNA linker, that provides covalent linkage between the first and second structures or domains. In embodiments, the RNA linker comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ribonucleotides. The TREM may comprise one or more linkers, for example, in embodiments, a TREM comprising (a), (b), (c), (d), and (e) may have a first linker between the first and second domains, and a second linker between the third domain and the further domain.

In embodiments, a TREM comprises an RNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical or differs by no more than 1, 2, 3, 4, 5, 10, 15, 20, 25, or 30 ribonucleotides from an RNA sequence encoded by a DNA sequence listed in table 1, or a fragment or functional fragment thereof. In embodiments, the TREM comprises an RNA sequence encoded by a DNA sequence listed in table 1, or a fragment or functional fragment thereof. In embodiments, the TREM comprises an RNA sequence encoded by a DNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence listed in table 1, or a fragment or functional fragment thereof. In embodiments, a TREM comprises a TREM domain, e.g., a domain described herein, that is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical or differs by no more than 1, 2, 3, 4, 5, 10, or 15 ribonucleotides, or a fragment or functional fragment thereof, to an RNA encoded by a DNA sequence listed in table 1. In embodiments, the TREM comprises a TREM domain, e.g., a domain described herein, comprising an RNA sequence encoded by a DNA sequence listed in table 1, or a fragment or functional fragment thereof. In embodiments, a TREM comprises a TREM domain, e.g., a domain described herein, comprising an RNA sequence encoded by a DNA sequence at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence listed in table 1, or a fragment or functional fragment thereof.

In embodiments, the TREM is 76-90 nucleotides in length. In embodiments, a TREM or fragment or functional fragment thereof is 10-90 nucleotides, 10-80 nucleotides, 10-70 nucleotides, 10-60 nucleotides, 10-50 nucleotides, 10-40 nucleotides, 10-30 nucleotides, 10-20 nucleotides, 20-90 nucleotides, 20-80 nucleotides, 20-70 nucleotides, 20-60 nucleotides, 20-50 nucleotides, 20-40 nucleotides, 30-90 nucleotides, 30-80 nucleotides, 30-70 nucleotides, 30-60 nucleotides, or 30-50 nucleotides.

In embodiments, TREM is aminoacylated, e.g., by charging an amino acid with an aminoacyl tRNA synthetase.

In embodiments, TREM is not loaded with an amino acid, e.g., TREM (utrem) unloaded.

In embodiments, the TREM comprises less than a full length tRNA. In embodiments, the TREM can correspond to a naturally occurring fragment of a tRNA, or to a non-naturally occurring fragment. Exemplary fragments include: TREM half (e.g., from a cleavage in ACHD, e.g., in an anticodon sequence, e.g., a 5 'half or a 3' half); a 5 'fragment (e.g., a fragment comprising the 5' terminus, e.g., from cleavage in DHD or ACHD); 3 'fragments (e.g., fragments comprising a 3' terminus, e.g., from cleavage in THD); or an internal fragment (e.g., a cut from one or more of ACHD, DHD, or THD).

The term "TREM composition" as used herein refers to a composition comprising a plurality of TREMs. A TREM composition may comprise one species or multiple species of TREM. In embodiments, the composition comprises only a single species of TREM. In embodiments, the TREM composition comprises a first TREM species and a second TREM species. In embodiments, the TREM composition comprises an X TREM species, wherein X ═ 2, 3, 4, 5, 6, 7, 8, 9, or 10. In embodiments, the TREM is at least 70%, 75%, 80%, 85%, 90%, or 95% identical, or 100% identical, to a sequence encoded by a nucleic acid in table 1. A TREM composition may comprise one species or multiple species of TREM. In embodiments, TREM compositions are purified from cell cultures. In embodiments, the cell culture from which TREM is purified comprises at least 1x 10⁷A hostCell, 1X 10⁸1X 10 host cell⁹1X 10 host cell¹⁰1X 10 host cell¹¹1X 10 host cell¹²1X 10 host cell¹³A host cell, or 1x 10¹⁴A host cell. In embodiments, the TREM composition is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% dry weight TREM (for a liquid composition, dry weight refers to removal of substantially all liquid, e.g., after lyophilization). In embodiments, the composition is a liquid. In embodiments, the composition is a dried, e.g., lyophilized, material. In an embodiment, the composition is a frozen composition. In embodiments, the composition is sterile. In embodiments, the composition comprises at least 0.5g, 1.0g, 5.0g, 10g, 15g, 25g, 50g, 100g, 200g, 400g, or 500g TREM (e.g., as determined by dry weight).

As used herein, "tumor suppressor" refers to a gene that modulates one or more cellular processes, including: cell fate determination, cell survival and genome maintenance. In embodiments, the tumor suppressor provides a selective growth advantage to a cell whose disorder (e.g., a genetic disorder (e.g., a mutation or deletion) or epigenetic disorder) is deregulated. Exemplary tumor suppressors include p53 or Rb.

Host cell

The host cell is a cell (e.g., a cultured cell) that can be used to express and/or purify a TREM. In embodiments, the host cell comprises a mammalian cell, e.g., a human cell. In embodiments, the host cell comprises a non-mammalian cell, e.g., a yeast cell. In embodiments, host cells include HeLa cells, HEK293T cells (e.g., Freestyle 293-F cells), HT-1080 cells, PER. C6 cells, HKB-11 cells, CAP cells, HuH-7 cells, BHK 21 cells, MRC-S cells, MDCK cells, VERO cells, WI-38 cells, or Chinese Hamster Ovary (CHO) cells. In embodiments, the host cell comprises a cancer cell, e.g., a solid tumor cell (e.g., a breast cancer cell (e.g., MCF7 cell), a pancreatic cell line (e.g., MIA PaCa-2 cell), a lung cancer cell, or a prostate cancer cell, or a blood cancer cell). In embodiments, the host cell comprises a cell that expresses one or more tissue-specific trnas. For example, a host cell can include a cell derived from a tissue associated with expression of a tRNA, e.g., a tissue-specific tRNA. In embodiments, a host cell expressing a tissue-specific tRNA is modified to express a TREM or fragment thereof.

In embodiments, the host cell is not a bacterial cell, such as an e.

In embodiments, the host cell is a cell that can be maintained under conditions that allow expression of TREM.

In embodiments, the host cell is capable of post-transcriptionally modifying TREM, e.g., adding a post-transcriptional modification selected from table 2. In the examples, the host cell expresses (e.g., naturally or heterologously) the enzymes listed in table 2. In embodiments, the host cell expresses (e.g., naturally or heterogeneously) an enzyme, e.g., an enzyme having nuclease activity (e.g., endonuclease activity or ribonuclease activity), e.g., or one or more of dicer, angiogenin, rnase A, RNA, enzyme P, RNA, enzyme Z, Rny1, or PrrC.

Method for culturing host cells

The host cell may be cultured in a medium that promotes growth (e.g., proliferation or hyperproliferation of the host cell). The host cell may be cultured in a suitable medium, for example, any of the following media: DMEM, MEM alpha, RPMI, F-10 medium, F-12 medium, DMEM/F-12 medium, IMDM, Medium 199, Leibovitz L-15, McCoys's 5A, MDCB medium, or CMRL medium. In an embodiment, the medium is supplemented with glutamine. In an embodiment, the medium is not supplemented with glutamine. In embodiments, the host cell is cultured in a medium with excess nutrients, e.g., not nutrient limiting. The host cell may be cultured in a medium comprising or supplemented with: one or a combination of growth factors, cytokines or hormones, for example, one or a combination of serum (e.g., Fetal Bovine Serum (FBS)), HEPES, Fibroblast Growth Factor (FGF), Epidermal Growth Factor (EGF), insulin-like growth factor (IGF), transforming growth factor beta (TGFb), platelet-derived growth factor (PDGF), Hepatocyte Growth Factor (HGF) or Tumor Necrosis Factor (TNF).

The host cell may also be cultured under conditions that induce stress, for example, cellular stress, osmotic stress, transformation stress, or oncogenic stress. In embodiments, a host cell expressing TREM is cultured under conditions that induce stress (e.g., as described herein) to produce a fragment of TREM, e.g., as described herein.

The host cell may be cultured under nutrient limiting conditions, e.g., the host cell is cultured in a medium having a limited amount of one or more nutrients. Examples of nutrients that may be limited are amino acids, lipids, carbohydrates, hormones, growth factors or vitamins. In embodiments, a host cell expressing TREM is cultured in a medium having a limiting amount of one or more nutrients, e.g., a nutrient deficiency of the medium, resulting in a fragment of TREM, e.g., as described herein. In embodiments, a host cell expressing TREM is cultured in a medium having a limiting amount of one or more nutrients, e.g., the medium is nutrient deficient, resulting in an unloaded TREM (e.g., uTREM).

The host cell may comprise an immortalized cell, e.g., a cell that expresses one or more enzymes involved in immortalization (e.g., TERT). In embodiments, the host cell may be propagated indefinitely.

The host cells may be cultured in suspension or as a monolayer. The host cell culture may be performed in a cell culture vessel or bioreactor. The cell culture vessel comprises a cell culture dish, plate, or flask. Exemplary cell culture vessels include 35mm, 60mm, 100mm or 150mm petri dishes, multi-well plates (e.g., 6-well, 12-well, 24-well, 48-well or 96-well plates), or T-25, T-75 or T-160 flasks.

In an embodiment, the host cell may be cultured in a bioreactor. The bioreactor may be, for example, a continuous flow batch bioreactor, a perfusion bioreactor, a batch process bioreactor or a fed-batch bioreactor. The bioreactor may be maintained under conditions sufficient to express TREM. Culture conditions can be adjusted to optimize the yield, purity or structure of TREM. In an embodiment, the bioreactor comprises1x 10 less⁷、1x 10⁸、1x 10⁹、1x 10¹⁰、1x 10¹¹、1x 10¹²、1x 10¹³Or 1x 10¹⁴A host cell. In an embodiment, the bioreactor comprises 1x 10⁷To 1x 10¹⁴A host cell; 1x 10⁷To 0.5x 10¹⁴A host cell; 1x 10⁷To 1x 10¹³A host cell; 1x 10⁷To 0.5x 10¹³A host cell; 1x 10⁷To 1x 10¹²A host cell; 1x 10⁷To 0.5x 10¹²A host cell; 1x 10⁷To 1x 10 ¹¹A host cell; 1x10⁷To 0.5x 10¹¹A host cell; 1x10⁷To 1x10¹⁰A host cell; 1x10⁷To 0.5x 10¹⁰A host cell; 1x10⁷To 1x10⁹A host cell; 1x10⁷To 0.5x 10⁹A host cell; 1x10⁷To 1x10⁸A host cell; 1x10⁷To 0.5x 10⁸A host cell; 0.5x 10⁸To 1x10¹⁴A host cell; 1x10⁸To 1x10¹⁴A host cell; 0.5x 10⁹To 1x10¹⁴A host cell; 1x10⁹To 1x10¹⁴A host cell; 0.5x 10¹⁰To 1x10¹⁴A host cell; 1x10¹⁰To 1x10¹⁴A host cell; 0.5x 10¹¹To 1x10¹⁴A host cell; 1x10¹¹To 1x10¹⁴A host cell; 0.5x 10¹²To 1x10¹⁴A host cell; 1x10¹²To 1x10¹⁴A host cell; 0.5x 10¹³To 1x10¹⁴A host cell; 1x10¹³To 1x10¹⁴A host cell; or 0.5x 10¹³To 1x10¹⁴A host cell.

In an embodiment, the bioreactor comprises at least 1x10⁵Individual host cell/mL, 2X 10⁵Individual host cell/mL, 3X 10⁵Individual host cell/mL, 4X 10⁵Individual host cell/mL, 5X 10⁵Individual host cell/mL, 6X 10⁵Individual host cell/mL, 7X 10⁵Individual host cell/mL, 8X 10⁵Individual host cell/mL, 9X 10⁵1X10 individual host cells/mL⁶Individual host cell/mL, 2X 10⁶Individual host cell/mL, 3X 10 ⁶Individual host cell/mL, 4X 10⁶Individual host cell/mL, 5X 10⁶Individual host cell/mL, 6X 10⁶Individual host cell/mL, 7X 10⁶Individual host cell/mL, 8X 10⁶Individual host cell/mL, 9X10⁶1X 10 individual host cells/mL⁷Individual host cell/mL, 2X 10⁷Individual host cell/mL, 3X 10⁷Individual host cell/mL, 4X 10⁷Individual host cell/mL, 5X 10⁷Individual host cell/mL, 6X 10⁷Individual host cell/mL, 7X 10⁷Individual host cell/mL, 8X 10⁷Individual host cell/mL, 9X10⁷1X 10 individual host cells/mL⁸Individual host cell/mL, 2X 10⁸Individual host cell/mL, 3X 10⁸Individual host cell/mL, 4X 10⁸Individual host cell/mL, 5X 10⁸Individual host cell/mL, 6X 10⁸Individual host cell/mL, 7X 10⁸Individual host cell/mL, 8X 10⁸Individual host cell/mL, 9X10⁸Individual host cell/mL, or 1X 10⁹Individual host cells/mL. In an embodiment, the bioreactor comprises 1x 10⁵Host cells/mL to 1x 10⁹Individual host cell/mL, 5X 10⁵Host cells/mL to 1x 10⁹1X 10 individual host cells/mL⁶Host cells/mL to 1x 10⁹Individual host cells/mL; 5x 10⁶Host cells/mL to 1x 10⁹1X 10 individual host cells/mL⁷Host cells/mL to 1x 10⁹Individual host cell/mL, 5X 10⁷Host cells/mL to 1x 10 ⁹1X10 individual host cells/mL⁸Host cells/mL to 1x10⁹Individual host cell/mL, 5X 10⁸Host cells/mL to 1x10⁹1X10 individual host cells/mL⁵Host cells/mL to 5X 10⁸1X10 individual host cells/mL⁵Host cells/mL to 1x10⁸1X10 individual host cells/mL⁵Per host cell/mL to5x 10⁷1X10 individual host cells/mL⁵Host cells/mL to 1x10⁷1X10 individual host cells/mL⁵Host cells/mL to 5X 10⁶1X10 individual host cells/mL⁵Host cells/mL to 1x10⁶Individual host cell/mL, or 1X10⁵Host cells/mL to 5X 10⁵Individual host cells/mL.

In an embodiment, the batch process bioreactor comprises 1x10⁶To 1x10⁷Individual host cells/ml.

In an embodiment, a batch process bioreactor having a volume of 100mL comprises 1x10⁸To 1x10⁹A host cell.

In an embodiment, a batch process bioreactor having a volume of 100L comprises 1x10¹¹To 1x10¹²A host cell.

In an embodiment, the fed batch bioreactor comprises 1x10⁷To 3x10⁷Individual host cells/ml.

In an example, a fed batch bioreactor with a volume of 100mL comprises 1x10⁹To 3x10⁹A host cell.

In an example, a fed batch bioreactor with a volume of 100L comprises 1x10 ¹²To 3x10¹²A host cell.

In an embodiment, the perfusion bioreactor comprises 1x 10⁸Individual host cells/ml.

In an embodiment, a perfusion bioreactor having a volume of 100mL comprises 1x 10¹⁰A host cell.

In an embodiment, a perfusion bioreactor having a volume of 100L comprises 1x 10¹³A host cell.

In embodiments, the bioreactor is maintained under conditions that promote growth of the host cells, e.g., at a temperature (e.g., 37 ℃) and a gas concentration (e.g., 5% CO) that allow growth of the host cells₂) The following steps.

For example, in some aspects, a bioreactor unit may perform one or more or all of the following: supply of nutrients and/or carbon sources, injection of suitable gases (e.g., oxygen), inlet and outlet flow of fermentation or cell culture media, separation of gas and liquid phases, maintenance of temperature, maintenance of oxygen and CO2 levels, maintenance of pH levels, agitation (e.g., stirring), and/or cleaning/disinfection. Exemplary bioreactor units, multiple reactors may be contained within a unit, for example the unit may have 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90 or 100 or more bioreactors in each unit and/or the facility may contain multiple units with single or multiple reactors within the facility. Any suitable bioreactor diameter may be used.

In embodiments, the volume of the bioreactor may be about 100mL to about 100L. Non-limiting examples include volumes of 100mL, 250mL, 500mL, 750mL, 1 liter, 2 liters, 3 liters, 4 liters, 5 liters, 6 liters, 7 liters, 8 liters, 9 liters, 10 liters, 15 liters, 20 liters, 25 liters, 30 liters, 40 liters, 50 liters, 60 liters, 70 liters, 80 liters, 90 liters, 100 liters. Further, suitable reactors may be multi-use, single-use, disposable, or non-disposable, and may be formed of any suitable material, including metal alloys, such as stainless steel (e.g., 316L or any other suitable stainless steel), and inconel, plastic, and/or glass. In some embodiments, a suitable reactor may be circular, e.g., cylindrical. In some embodiments, a suitable reactor may be square, e.g., rectangular. In certain instances, a square reactor may provide benefits over a round reactor, such as ease of use (e.g., loading and setup by a technician), better mixing and uniformity of the reactor contents, and a lower footprint.

Method for modifying host cells

The host cell can be modified to optimize production of TREM, e.g., to have optimized TREM yield, purity, structure (e.g., folding), or stability. In embodiments, the host cell can be modified (e.g., using the methods described herein) to increase or decrease expression of a desired molecule (e.g., a gene that optimizes TREM production (e.g., optimizes yield, purity, structure, or stability of TREM). In embodiments, the host cell may be epigenetically modified, e.g., using the methods described herein, to increase or decrease expression of a desired gene (which optimizes production).

In embodiments, the host cell can be modified to increase or decrease expression of an oncogene (e.g., as described herein), a tumor suppressor (e.g., as described herein), or a molecule involved in tRNA or TREM modulation (e.g., a gene involved in tRNA or TREM transcription, processing, modification, stability, or folding). Exemplary oncogenes include Myc (e.g., c-Myc, N-Myc or L-Myc), c-Jun, Wnt or RAS. Exemplary tumor suppressors include p53 or Rb. Exemplary molecules involved in tRNA or TREM modulation include: RNA polymerase III (Pol III) and Pol III accessory molecules (e.g., TFIIIB); maf1, Trm1, Mck1, or Kns 1; enzymes involved in tRNA or TREM modification, e.g., genes listed in Table 2; or a molecule having nuclease activity, e.g., or one or more of dicer, angiogenin, rnase A, RNA, P, RNA, enzyme Z, Rny1, or PrrC.

In embodiments, the host cell may be modified by: transfection (e.g., transient transfection or stable transfection); transduction (e.g., viral transduction, e.g., lentiviral, adenoviral or retroviral transduction); electroporation; lipid-based agent delivery (e.g., liposomes), nanoparticle-based agent delivery; or other methods known in the art.

In embodiments, the host cell may be modified to increase expression, e.g., overexpression, of: desired molecules, e.g., genes (e.g., oncogenes, or genes involved in tRNA or TREM modulation (e.g., genes encoding the enzymes listed in table 2, or genes encoding enzymes having nuclease activity (e.g., endonuclease activity or ribonuclease activity), e.g., or one or more of dicer, angiogenin, rnase A, RNA enzyme P, RNA enzyme Z, Rny1, or PrrC exemplary methods of increasing gene expression include (a) contacting a host cell with a nucleic acid (e.g., DNA or RNA) encoding a gene, (b) contacting a host cell with a peptide that expresses a target protein, (c) contacting a host cell with a molecule that modulates, e.g., increases expression of a target gene (e.g., a small RNA (e.g., microrna or small interfering RNA) or a low molecular weight compound), or (d) contacting a host cell with a gene editing moiety (e.g., Zinc Finger Nuclease (ZFN) or zinc finger 9/CRISPR molecule), the gene editing portion inhibits (e.g., mutates or knocks out) the expression of a target gene negative regulator. In embodiments, a nucleic acid encoding a gene or a plasmid containing a nucleic acid encoding a gene can be introduced into a host cell by transfection or electroporation. In embodiments, a nucleic acid encoding a gene can be introduced into a host cell by contacting the host cell with a virus (e.g., a lentivirus, adenovirus, or retrovirus) that expresses the gene.

In embodiments, the host cell can be modified to reduce, e.g., minimize, the expression of a desired molecule, e.g., a gene (e.g., a tumor suppressor gene, or a gene involved in tRNA or TREM regulation). Exemplary methods of reducing gene expression include: (a) contacting a host cell with a nucleic acid (e.g., DNA or RNA) encoding a gene inhibitor (e.g., a gene or a dominant negative variant or negative regulator of a protein encoded by the gene); (b) contacting a host cell with a peptide that inhibits a target protein; (c) contacting a host cell with a molecule that modulates, e.g., inhibits, the expression of a target gene (e.g., a small RNA (e.g., a microrna or a small interfering RNA) or a low molecular weight compound); or (d) contacting the host cell with a gene editing moiety, such as a Zinc Finger Nuclease (ZFN) or Cas9/CRISPR molecule, that inhibits (e.g., mutates or knocks out) expression of the target gene. In embodiments, a nucleic acid encoding a gene inhibitor or a plasmid containing a nucleic acid encoding a gene inhibitor can be introduced into a host cell by transfection or electroporation. In embodiments, a nucleic acid encoding a gene inhibitor can be introduced into a host cell by contacting the host cell with a virus that expresses the gene inhibitor (e.g., a lentivirus, adenovirus, or retrovirus).

In embodiments, a host cell (e.g., a host cell described herein) is modified (e.g., by transfection with a nucleic acid) to express, e.g., overexpress, an oncogene, e.g., a c-Myc, described herein.

In embodiments, a host cell (e.g., a host cell described herein) is modified (e.g., by transfection with a nucleic acid) to inhibit, e.g., down-regulate, expression of a tumor suppressor, e.g., a tumor suppressor described herein, e.g., p53 or Rb.

In embodiments, a host cell (e.g., a HEK293T cell) is modified (e.g., using a CRISPR/Cas9 molecule) to inhibit, e.g., knock out, expression of a gene (e.g., Maf1) that modulates a tRNA or TREM. In embodiments, a host cell (e.g., a HEK293T cell) is modified to overexpress a gene that modulates a tRNA or TREM, e.g., Trm 1.

In embodiments, a host cell (e.g., a HEK293T cell) is modified to overexpress a gene that modulates a tRNA or TREM (e.g., Trm1) and to overexpress an oncogene (e.g., an oncogene described herein, e.g., c-Myc).

TREM

A "tRNA based effector molecule" or "TREM" refers to an RNA molecule that comprises one or more of the properties described herein. TREM can be loaded with amino acids, e.g., homologous amino acids; non-homologous amino acids are loaded (e.g., erroneously loaded TREM (mTREM); or unloaded, e.g., unloaded TREM (uTREM)).

In embodiments, a TREM comprises a ribonucleic acid (RNA) sequence encoded by a deoxyribonucleic acid (DNA) sequence disclosed in Table 1 (e.g., any one of SEQ ID NOS: 1-451 disclosed in Table 1). In embodiments, a TREM comprises an RNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence disclosed in table 1 (e.g., any one of SEQ ID NOs 1-451 disclosed in table 1). In embodiments, a TREM comprises an RNA sequence encoded by a DNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in table 1 (e.g., any one of SEQ ID NOs 1-451 disclosed in table 1).

In embodiments, a TREM comprises at least 30 contiguous nucleotides of an RNA sequence encoded by a DNA sequence disclosed in table 1, e.g., at least 30 contiguous nucleotides of an RNA sequence encoded by any one of SEQ ID NOs 1-451 disclosed in table 1. In embodiments, a TREM comprises at least 30 contiguous nucleotides of an RNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence disclosed in table 1 (e.g., any one of SEQ ID NOs 1-451 disclosed in table 1). In embodiments, a TREM comprises at least 30 contiguous nucleotides of an RNA sequence encoded by a DNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in table 1 (e.g., any one of SEQ ID NOs 1-451 disclosed in table 1).

In embodiments, a TREM, e.g., an exogenous TREM, comprises 1, 2, 3, or 4 of the following properties:

(a) at least one nucleotide or one post-transcriptional modification that differs from the closest sequence tRNA in a reference cell, e.g., a cell into which an exogenous nucleic acid is introduced;

(b) cells other than the cells into which they have been introduced for transcription;

(d) Has an expression profile, e.g., level or profile, which is not wild-type, e.g., its expression level is higher than wild-type.

In embodiments, expression profiling can be mediated by introducing changes into nucleic acids that modulate expression or by adding agents that modulate expression of RNA molecules.

In embodiments, a TREM, e.g., an exogenous TREM, includes (a), (b), (c), and (d).

In embodiments, a TREM, e.g., an exogenous TREM, includes (a), (b), and (c).

In embodiments, a TREM, e.g., an exogenous TREM, includes (a), (b), and (d).

In embodiments, a TREM, e.g., an exogenous TREM, includes (a), (c), and (d).

In embodiments, a TREM, e.g., an exogenous TREM, includes (b), (c), and (d).

In embodiments, a TREM, e.g., an exogenous TREM, comprises (a) and (d).

In embodiments, a TREM, e.g., an exogenous TREM, comprises (c) and (d).

TREM fragment

In embodiments, a TREM comprises a fragment (sometimes referred to herein as a TREM fragment), e.g., a fragment of an RNA encoded by a deoxyribonucleic acid sequence disclosed in table 1. For example, a TREM includes less than the entire sequence of a tRNA from the same species as the subject being treated, e.g., less than the entire sequence of a tRNA having the same anticodon, or both. In embodiments, production of TREM fragments, e.g., from full-length TREM or longer fragments, can be catalyzed by an enzyme, e.g., an enzyme having nuclease activity (e.g., endonuclease activity or ribonuclease activity), such as dicer, angiogenin, rnase P, RNA enzyme Z, Rny1, or PrrC.

In embodiments, TREM fragments can be generated in vivo, ex vivo, or in vitro. In embodiments, the TREM fragment is produced in vivo in a host cell. In embodiments, the TREM fragment is generated ex vivo. In embodiments, TREM fragments are generated in vitro, as described in example 12. In embodiments, the TREM fragment is produced by fragmenting an expressed TREM after the cell produces a TREM, e.g., the TREM produced by the host cell is fragmented after release or purification from the host cell, e.g., the TREM is fragmented ex vivo in vitro.

Exemplary TREM fragments include a TREM half (e.g., from cleavage in ACHD, e.g., a 5 'TREM half or a 3' TREM half), a 5 'fragment (e.g., a fragment comprising a 5' terminus, e.g., from cleavage in DHD or ACHD), a 3 'fragment (e.g., a fragment comprising a 3' terminus of TREM, e.g., from cleavage in THD), or an internal fragment (e.g., from cleavage in one or more of ACHD, DHD, or THD).

In embodiments, a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% of the RNA sequence encoded by a DNA sequence provided in table 1 (e.g., any one of SEQ ID NOs: 1-451 disclosed in table 1). In embodiments, a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% of an RNA sequence that is at least 80%, 85%, 90%, 95%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence provided in table 1 (e.g., any one of SEQ ID NOs 1-451 disclosed in table 1). In embodiments, a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% of an RNA sequence encoded by a DNA sequence that is at least 80%, 85%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in table 1 (e.g., any one of SEQ ID NOs 1-451 disclosed in table 1).

In embodiments, a TREM fragment comprises at least 5 ribonucleotides (nt), 10nt, 15nt, 20nt, 25nt, 30nt, 35nt, 40nt, 45nt, 50nt, 55nt, or 60nt (but less than full-length) of an RNA sequence encoded by a DNA sequence disclosed in table 1 (e.g., any one of SEQ ID NOs: 1-451 disclosed in table 1). In embodiments, a TREM fragment comprises at least 5 ribonucleotides (nt), 10nt, 15nt, 20nt, 25nt, 30nt, 35nt, 40nt, 45nt, 50nt, 55nt, or 60nt (but less than full length) of an RNA sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an RNA sequence encoded by a DNA sequence provided in table 1 (e.g., any one of SEQ ID NOs: 1-451 disclosed in table 1). In embodiments, a TREM fragment comprises at least 5 ribonucleotides (nt), 10nt, 15nt, 20nt, 25nt, 30nt, 35nt, 40nt, 45nt, 50nt, 55nt, or 60nt (but less than full length) of an RNA sequence encoded by a DNA sequence having at least 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, or 100% identity to a DNA sequence provided in table 1 (e.g., any of SEQ ID NOs: 1-451 disclosed in table 1).

In embodiments, a TREM fragment comprises a sequence 10-90 ribonucleotides (rnt), 10-80rnt, 10-70rnt, 10-60rnt, 10-50rnt, 10-40rnt, 10-30rnt, 10-20rnt, 20-90rnt, 20-80rnt, 20-70rnt, 20-60rnt, 20-50rnt, 20-40rnt, 30-90rnt, 30-80rnt, 30-70rnt, 30-60rnt, or 30-50rnt in length.

In embodiments, a TREM fragment comprises a TREM structure, domain, or activity, e.g., as described above. In embodiments, TREM fragments comprise adaptor functions, e.g., as described herein. In embodiments, TREM fragments comprise homologous adaptor functions, e.g., as described herein. In embodiments, TREM fragments comprise non-homologous adaptor functions, e.g., as described herein. In embodiments, the TREM fragment comprises a regulatory function, e.g., as described herein.

In embodiments, the TREM fragment comprises a translation-inhibitory function, e.g., a shift in an initiation factor such as eIF 4G.

In embodiments, TREM fragments comprise epigenetic functions, e.g., epigenetics of a disorder, e.g., a metabolic disorder. In some embodiments, the epigenetic function may have a generational impact, e.g., as compared to somatic epigenetic regulation.

In embodiments, a TREM fragment comprises a retroviral regulatory function, e.g., modulation of retroviral reverse transcription, e.g., HERV regulation.

In embodiments, a TREM fragment comprises a gene silencing function, e.g., by binding to an AGO and/or PIWI.

In embodiments, a TREM fragment comprises a neuroprotective function, e.g., by chelating translation initiation factors, e.g., in a stress particle, to promote survival of a motor neuron, e.g., under cellular stress.

In embodiments, TREM fragments comprise an anti-cancer function, e.g., preventing cancer progression by binding and/or sequestering, e.g., a metastatic transcript stabilizing protein.

In embodiments, TREM fragments comprise a cell survival function, e.g., increasing cell survival by binding to, e.g., cytochrome c and/or cyt c ribonucleoprotein complexes.

In embodiments, TREM fragments comprise a ribosomal biogenesis function, e.g., TREM fragments can modulate ribosomal biogenesis by, e.g., modulating or binding to mRNA encoding a ribosomal protein.

TREM modification

The TREMs described herein can comprise a moiety, generally referred to herein as a modification, e.g., a moiety described in table 2. Although the term modification as used herein should not generally be construed as a product of any particular process, in embodiments, the formation of the modification may be mediated by the enzymes in table 2. In embodiments, the modification is post-transcriptional. In embodiments, the modification is co-transcription formation. In embodiments, the modification occurs in vivo, e.g., in a host cell.

In embodiments, the modification is a modification listed in any one of lines 1-62 of table 2. In embodiments, the modification is a modification set forth in any one of rows 1-62 of table 2, and the formation of the modification is mediated by an enzyme in table 2. In embodiments, the modifications are selected from the rows in table 2 and the formation of the modifications is mediated by enzymes from the same rows in table 2.

Table 2: list of tRNA modifications and related enzymes.

TREM fusion

In embodiments, the TREMs disclosed herein comprise additional moieties, e.g., fusion moieties. In embodiments, the fusion moiety can be used for purification to alter the folding of TREM or as a targeting moiety. In embodiments, the fusion moiety may comprise a tag, linker, may be cleavable, or may comprise a binding site for an enzyme. In embodiments, the fusion moiety can be located at the N-terminus of TREM or the C-terminus of TREM. In embodiments, the fusion moiety can be encoded by the same or different nucleic acid molecule encoding TREM.

TREM consensus sequence

In embodiments, the TREMs disclosed herein comprise a consensus sequence provided herein.

In embodiments, a TREM disclosed herein comprises formula I_ZZZWherein_ZZZRepresents any of the twenty amino acids and formula I corresponds to all species.

In embodiments, a TREM disclosed herein comprises formula II_ZZZOf the consensus sequence of (1), wherein_ZZZRepresents any of the twenty amino acids and formula II corresponds to a mammal.

In embodiments, a TREM disclosed herein comprises formula III_ZZZOf the consensus sequence of (1), wherein_ZZZRepresents any of the twenty amino acids and formula III corresponds to human.

In an embodiment of the present invention,_ZZZrepresents any of twenty amino acids: alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine or valine.

In embodiments, the TREM disclosed herein comprises a property selected from the group consisting of:

a) under physiological conditions, the residue R₀Forming a linker region, e.g., linker 1 region;

b) residue R under physiological conditions₁-R₂-R₃-R₄-R₅-R₆-R₇And a residue R₆₅-R₆₆-R₆₇-R₆₈-R₆₉-R₇₀-R₇₁Forming a stem region, e.g., an AStD stem region;

c) under physiological conditions, the residue R₈-R₉Forming a linker region, e.g., linker 2 region;

e) residue-R under physiological conditions₂₉Forming a linker region, e.g., linker 3 region;

g) residue- [ R ] under physiological conditions₄₇]_xComprises a variable region, e.g., as described herein;

i) Residue R under physiological conditions₇₂A tab region, e.g., tab 4 region, is formed.

Alanine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_ALAThe sequence of (SEQ ID NO:562),

wherein R is a ribonucleotide residue and is common to Ala:

R₀absent;

R₁₄、R₅₇independently is a or absent;

R₂₆a, C, G or absent;

R₅、R₆、R₁₅、R₁₆、R₂₁、R₃₀、R₃₁、R₃₂、R₃₄、R₃₇、R₄₁、R₄₂、R₄₃、R₄₄、R₄₅、R₄₈、R₄₉、R₅₀、R₅₈、R₅₉、R₆₃、R₆₄、R₆₆、R₆₇independently N or absent;

R₁₁、R₃₅、R₆₅independently A, C, U or absent;

R₁、R₉、R₂₀、R₃₈、R₄₀、R₅₁、R₅₂、R₅₆independently A, G or absent;

R₇、R₂₂、R₂₅、R₂₇、R₂₉、R₄₆、R₅₃、R₇₂independently A, G, U or absent;

R₂₄、R₆₉independently A, U or absent;

R₇₀、R₇₁independently is C or absent;

R₃、R₄independently C, G or absent;

R₁₂、R₃₃、R₃₆、R₆₂、R₆₈independently C, G, U or absent;

R₁₃、R₁₇、R₂₈、R₃₉、R₅₅、R₆₀、R₆₁independently C, U or absent;

R₁₀、R₁₉、R₂₃independently G or absent;

R₂g, U or absent;

R₈、R₁₈、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

in this case, for example, x is 1 to 271 (e.g., x is 1 to 250, x is 1 to 225, x is 1 to 200, x is 1 to 175, x is 1 to 150, x is 1 to 125, x is 1 to 100, x is 1 to 75, x is 1 to 50, x is 1 to 40, x is 1 to 30, x is 1 to 29, x is 1 to 28, x is 1 to 27, x is 1 to 26, x is 1 to 25, x is 1 to 24, x is 1 to 23, x is 1 to 22, x is 1 to 21, x is 1 to 20, x is 1 to 19, x is 1 to 18, x is 1 to 17, x is 1 to 16, x is 1 to 15, x is 1 to 14, x is 1 to 13, x is 1 to 17, x is 1 to 16, x is 1 to 15, x is 1 to 14, x is 1 to 12, x is 1 to 13, x-1-10, x-10-271, x-20-271, x-30-271, x-40-271, x-50-271, x-60-271, x-70-271, x-80-271, x-100-271, x-125-271, x-150-271, x-175-271, x-200-271, x-225-271, x-1, x-2, x-3, x-4, x-5, x-6, x-7, x-8, x-9, x-10, x-11, x-12, x-13, x-14, x-15, x-16, x-20, x-30, x-20, x-30, x-20, x-271, x-20, x-15, x 90, x 100, x 110, x 125, x 150, x 175, x 200, x 225, x 250, or x 271),

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_ALAThe sequence of (SEQ ID NO:563),

wherein R is a ribonucleotide residue and is common to Ala:

R₀、R₁₈absent;

R₁₄、R₂₄、R₅₇independently is a or absent;

R₁₅、R₂₆、R₆₄independently A, C, G or absent;

R₁₆、R₃₁、R₅₀、R₅₉independently N or absent;

R₁₁、R₃₂、R₃₇、R₄₁、R₄₃、R₄₅、R₄₉、R₆₅、R₆₆independently A, C, U or absent;

R₁、R₅、R₉、R₂₅、R₂₇、R₃₈、R₄₀、R₄₆、R₅₁、R₅₆independently A, G or absent;

R₇、R₂₂、R₂₉、R₄₂、R₄₄、R₅₃、R₆₃、R₇₂independently A, G, U or absent;

R₆、R₃₅、R₆₉independently A, U or absent;

R₅₅、R₆₀、R₇₀、R₇₁independently is C or absent;

R₃c, G or absent;

R₁₂、R₃₆、R₄₈independently C, G, U or absent;

R₁₃、R₁₇、R₂₈、R₃₀、R₃₄、R₃₉、R₅₈、R₆₁、R₆₂、R₆₇、R₆₈independently C, U or absent;

R₄、R₁₀、R₁₉、R₂₀、R₂₃、R₅₂independently G or absent;

R₂、R₈、R₃₃independently G, U or absent;

R₂₁、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_ALAThe sequence of (SEQ ID NO:564),

wherein R is a ribonucleotide residue and is common to Ala:

R₀、R₁₈absent;

R₁₄、R₂₄、R₅₇、R₇₂independently is a or absent;

R₁₅、R₂₆、R₆₄independently A, C, G or absent;

R₁₆、R₃₁、R₅₀independently N or absent;

R₁₁、R₃₂、R₃₇、R₄₁、R₄₃、R₄₅、R₄₉、R₆₅、R₆₆independently A, C, U or absent;

R₅、R₉、R₂₅、R₂₇、R₃₈、R₄₀、R₄₆、R₅₁、R₅₆independently A, G or absent;

R₇、R₂₂、R₂₉、R₄₂、R₄₄、R₅₃、R₆₃independently A, G, U or absent;

R₆、R₃₅independently isA. U or absent;

R₅₅、R₆₀、R₆₁、R₇₀、R₇₁independently is C or absent;

R₁₂、R₄₈、R₅₉independently C, G, U or absent;

R₁₃、R₁₇、R₂₈、R₃₀、R₃₄、R₃₉、R₅₈、R₆₂、R₆₇、R₆₈independently C, U or absent;

R₁、R₂、R₃、R₄、R₁₀、R₁₉、R₂₀、R₂₃、R₅₂independently G or absent;

R₃₃、R₃₆independently G, U or absent;

R₈、R₂₁、R₅₄、R₆₉independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Arginine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_ARGThe sequence of (SEQ ID NO:565),

wherein R is a ribonucleotide residue and is common to Arg:

R₅₇a or absent;

R₉、R₂₇independently A, C, G or absent;

R₁、R₂、R₃、R₄、R₅、R₆、R₇、R₁₁、R₁₂、R₁₆、R₂₁、R₂₂、R₂₃、R₂₅、R₂₆、R₂₉、R₃₀、R₃₁、R₃₂、R₃₃、R₃₄、R₃₇、R₄₂、R₄₄、R₄₅、R₄₆、R₄₈、R₄₉、R₅₀、R₅₁、R₅₈、R₆₂、R₆₃、R₆₄、R₆₅、R₆₆、R₆₇、R₆₈、R₆₉、R₇₀、R₇₁independently N or absent;

R₁₃、R₁₇、R₄₁independently A, C, U or absent;

R₁₉、R₂₀、R₂₄、R₄₀、R₅₆independently A, G or absent;

R₁₄、R₁₅、R₇₂independently A, G, U or absent;

R₁₈a, U or absent;

R₃₈c or absent;

R₃₅、R₄₃、R₆₁independently C, G, U or absent;

R₂₈、R₅₅、R₅₉、R₆₀independently C, U or absent;

R₀、R₁₀、R₅₂independently G or absent;

R₈、R₃₉independently G, U or absent;

R₃₆、R₅₃、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_ARGThe sequence of (SEQ ID NO:566),

wherein R is a ribonucleotide residue and is common to Arg:

R₁₈absent;

R₂₄、R₅₇independently is a or absent;

R₄₁a, C or absent;

R₃、R₇、R₃₄、R₅₀independently A, C, G or absent;

R₂、R₅、R₆、R₁₂、R₂₆、R₃₂、R₃₇、R₄₄、R₅₈、R₆₆、R₆₇、R₆₈、R₇₀independently N or absent;

R₄₉、R₇₁independently A, C, U or absent;

R₁、R₁₅、R₁₉、R₂₅、R₂₇、R₄₀、R₄₅、R₄₆、R₅₆、R₇₂independently A, G or absent;

R₁₄、R₂₉、R₆₃independently A, G, U or absent;

R₁₆、R₂₁independently A, U or absent;

R₃₈、R₆₁independently is C or absent;

R₃₃、R₄₈independently C, G or absent;

R₄、R₉、R₁₁、R₄₃、R₆₂、R₆₄、R₆₉independently C, G, U or absent;

R₁₃、R₂₂、R₂₈、R₃₀、R₃₁、R₃₅、R₅₅、R₆₀、R₆₅independently C, U or absent;

R₀、R₁₀、R₂₀、R₂₃、R₅₁、R₅₂independently G or absent;

R₈、R₃₉、R₄₂independently G, U or absent;

R₁₇、R₃₆、R₅₃、R₅₄、R₅₉independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_ARGThe sequence of (SEQ ID NO:567),

R₀-R₁-R₂-R₃-R₄-R₅-R₆-R₇-R₈-R₉-R₁₀-R₁₁-R₁₂-R₁₃-R₁₄-R₁₅-R₁₆-R₁₇-R₁₈-R₁₉-R₂₀-R₂₁-R₂₂-R₂₃-R₂₄-R₂₅-R₂₆-R₂₇-R₂₈-R₂₉-R₃₀-R₃₁-R₃₂-R₃₃-R₃₄-R₃₅-R₃₆-R₃₇-R₃₈-R₃₉-R₄₀-R₄₁-R₄₂-R₄₃-R₄₄-R₄₅-R₄₆-[R₄₇]_x-R₄₈-R₄₉-R₅₀-R₅₁-R₅₂-R₅₃-R₅₄-R₅₅-R₅₆-_R57-R₅₈-R₅₉-R₆₀-R₆₁-R₆₂-R₆₃-R₆₄-R₆₅-R₆₆-R₆₇-R₆₈-R₆₉-R₇₀-R₇₁-R₇₂

wherein R is a ribonucleotide residue and is common to Arg:

R₁₈absent;

R₁₅、R₂₁、R₂₄、R₄₁、R₅₇independently is a or absent;

R₃₄、R₄₄independently A, C or absent;

R_3、R₅、R₅₈independently A, C, G or absent;

R₂、R₆、R₆₆、R₇₀independently N or absent;

R₃₇、R₄₉independently A, C, U or absent;

R₁、R₂₅、R₂₉、R₄₀、R₄₅、R₄₆、R₅₀independently A, G or absent;

R₁₄、R₆₃、R₆₈independently A, G, U or absent;

R₁₆a, U or absent;

R₃₈、R₆₁independently is C or absent;

R₇、R₁₁、R₁₂、R₂₆、R₄₈independently C, G or absent;

R₆₄、R₆₇、R₆₉independently C, G, U or absent;

R₄、R₁₃、R₂₂、R₂₈、R₃₀、R₃₁、R₃₅、R₄₃、R₅₅、R₆₀、R₆₂、R₆₅、R₇₁independently C, U or absent;

R₀、R₁₀、R₁₉、R₂₀、R₂₃、R₂₇、R₃₃、R₅₁、R₅₂、R₅₆、R₇₂independently G or absent;

R₈、R₉、R₃₂、R₃₉、R₄₂independently G, U or absent;

R₁₇、R₃₆、R₅₃、R₅₄、R₅₉independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Asparagine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_ASNThe sequence of (SEQ ID NO:568),

wherein R is a ribonucleotide residue and shared for Asn are:

R₀、R₁₈absent;

R₄₁a or absent;

R₁₄、R₄₈、R₅₆independently A, C, G or not(ii) present;

R₂、R₄、R₅、R₆、R₁₂、R₁₇、R₂₆、R₂₉、R₃₀、R₃₁、R₄₄、R₄₅、R₄₆、R₄₉、R₅₀、R₅₈、R₆₂、R₆₃、R₆₅、R₆₆、R₆₇、R₆₈、R₇₀、R₇₁independently N or absent;

R₁₁、R₁₃、R₂₂、R₄₂、R₅₅、R₅₉independently A, C, U or absent;

R₉、R₁₅、R₂₄、R₂₇、R₃₄、R₃₇、R₅₁、R₇₂independently A, G or absent;

R₁、R₇、R₂₅、R₆₉independently A, G, U or absent;

R₄₀、R₅₇independently A, U or absent;

R₆₀c or absent;

R₃₃c, G or absent;

R₂₁、R₃₂、R₄₃、R₆₄independently C, G, U or absent;

R₃、R₁₆、R₂₈、R₃₅、R₃₆、R₆₁independently C, U or absent;

R₁₀、R₁₉、R₂₀、R₅₂independently G or absent;

R₅₄g, U or absent;

R₈、R₂₃、R₃₈、R₃₉、R₅₃independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_ASNThe sequence of (SEQ ID NO:569),

wherein R is a ribonucleotide residue and shared for Asn are:

R_0、R₁₈is absent

R₂₄、R₄₁、R₄₆、R₆₂Independently is a or absent;

R₅₉a, C or absent;

R₁₄、R₅₆、R₆₆independently A, C, G or absent;

R₁₇、R₂₉independently N or absent;

R₁₁、R₂₆、R₄₂、R₅₅independently A, C, U or absent;

R₁、R₉、R₁₂、R₁₅、R₂₅、R₃₄、R₃₇、R₄₈、R₅₁、R₆₇、R₆₈、R₆₉、R₇₀、R₇₂independently A, G or absent;

R₄₄、R₄₅、R₅₈independently A, G, U or absent;

R₄₀、R₅₇independently A, U or absent;

R₅、R₂₈、R₆₀independently is C or absent;

R₃₃、R₆₅independently C, G or absent;

R₂₁、R₄₃、R₇₁independently C, G, U or absent;

R₃、R₆、R₁₃、R₂₂、R₃₂、R₃₅、R₃₆、R₆₁、R₆₃、R₆₄independently C, U or absent;

R₇、R₁₀、R₁₉、R₂₀、R₂₇、R₄₉、R₅₂independently G or absent;

R₅₄g, U or absent;

R₂、R₄、R₈、R₁₆、R₂₃、R₃₀、R₃₁、R₃₈、R₃₉、R₅₀、R₅₃independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_ASNThe sequence of (SEQ ID NO:570),

wherein R is a ribonucleotide residue and shared for Asn are:

R_0、R₁₈is absent

R₂₄、R₄₀、R₄₁、R₄₆、R₆₂Independently is a or absent;

R₅₉a, C or absent;

R₁₄、R₅₆、R₆₆independently A, C, G or absent;

R₁₁、R₂₆、R₄₂、R₅₅independently A, C, U or absent;

R₁、R₉、R₁₂、R₁₅、R₃₄、R₃₇、R₄₈、R₅₁、R₆₇、R₆₈、R₆₉、R₇₀independently A, G or absent;

R₄₄、R₄₅、R₅₈independently A, G, U or absent;

R₅₇a, U or absent;

R₅、R₂₈、R₆₀independently is C or absent;

R₃₃、R₆₅independently C, G or absent;

R₁₇、R₂₁、R₂₉independently C, G, U or absent;

R₃、R₆、R₁₃、R₂₂、R₃₂、R₃₅、R₃₆、R₄₃、R₆₁、R₆₃、R₆₄、R₇₁independently C, U or absent;

R₇、R₁₀、R₁₉、R₂₀、R₂₅、R₂₇、R₄₉、R₅₂、R₇₂independently G or absent;

R₅₄g, U or absent;

R₂、R₄、R₈、R₁₆、R₂₃、R₃₀、R₃₁、R₃₈、R₃₉、R₅₀、R₅₃independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Aspartic acid TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_ASPThe sequence of (SEQ ID NO:571),

wherein R is a ribonucleotide residue and common to Asp is:

R₀is absent

R₂₄、R₇₁Independently A, C or absent;

R₃₃、R₄₆independently A, C, G or absent;

R₂、R₃、R₄、R₅、R₆、R₁₂、R₁₆、R₂₂、R₂₆、R₂₉、R₃₁、R₃₂、R₄₄、R₄₈、R₄₉、R₅₈、R₆₃、R₆₄、R₆₆、R₆₇、R₆₈、R₆₉independently N or absent;

R₁₃、R₂₁、R₃₄、R₄₁、R₅₇、R₆₅independently A, C, U or absent;

R₉、R₁₀、R₁₄、R₁₅、R₂₀、R₂₇、R₃₇、R₄₀、R₅₁、R₅₆、R₇₂independently A, G or absent;

R₇、R₂₅、R₄₂independently A, G, U or absent;

R₃₉c or absent;

R₅₀、R₆₂independently C, G or absent;

R₃₀、R₄₃、R₄₅、R₅₅、R₇₀independently C, G, U or absent;

R₈、R₁₁、R₁₇、R₁₈、R₂₈、R₃₅、R₅₃、R₅₉、R₆₀、R₆₁independently C, U or absent;

R₁₉、R₅₂independently G or absent;

R₁g, U or absent;

R₂₃、R₃₆、R₃₈、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_ASPThe sequence of (SEQ ID NO:572),

wherein R is a ribonucleotide residue and common to Asp is:

R₀、R₁₇、R₁₈、R₂₃independently is absent;

R₉、R₄₀independently is a or absent;

R₂₄、R₇₁independently A, C or absent;

R₆₇、R₆₈independently A, C, G or absent;

R₂、R₆、R₆₆independently N or absent;

R₅₇、R₆₃independently A, C, U or absent;

R₁₀、R₁₄、R₂₇、R₃₃、R₃₇、R₄₄、R₄₆、R₅₁、R₅₆、R₆₄、R₇₂independently A, G or absent;

R₇、R₁₂、R₂₆、R₆₅independently A, U or absent;

R₃₉、R₆₁、R₆₂independently is C or absent;

R₃、R₃₁、R₄₅、R₇₀independently C, G or absent;

R₄、R₅、R₂₉、R₄₃、R₅₅independently C, G, U or absent;

R₈、R₁₁、R₁₃、R₃₀、R₃₂、R₃₄、R₃₅、R₄₁、R₄₈、R₅₃、R₅₉、R₆₀independently C, U or absent;

R₁₅、R₁₉、R₂₀、R₂₅、R₄₂、R₅₀、R₅₂independently G or absent;

R₁、R₂₂、R₄₉、R₅₈、R₆₉independently G, U or absent;

R₁₆、R₂₁、R₂₈、R₃₆、R₃₈、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In the examples, thisThe TREM disclosed herein comprises formula III_ASPThe sequence of (SEQ ID NO:573),

wherein R is a ribonucleotide residue and common to Asp is:

R₀、R₁₇、R₁₈、R₂₃is absent

R₉、R₁₂、R₄₀、R₆₅、R₇₁Independently is a or absent;

R₂、R₂₄、R₅₇independently A, C or absent;

R₆、R₁₄、R₂₇、R₄₆、R₅₁、R₅₆、R₆₄、R₆₇、R₆₈independently A, G or absent;

R₃、R₃₁、R₃₅、R₃₉、R₆₁、R₆₂independently is COr is absent;

R₆₆c, G or absent;

R₅、R₈、R₂₉、R₃₀、R₃₂、R₃₄、R₄₁、R₄₃、R₄₈、R₅₅、R₅₉、R₆₀、R₆₃independently C, U or absent;

R₁₀、R₁₅、R₁₉、R₂₀、R₂₅、R₃₃、R₃₇、R₄₂、R₄₄、R₄₅、R₄₉、R₅₀、R₅₂、R₆₉、R₇₀、R₇₂independently G or absent;

R₂₂、R₅₈independently G, U or absent;

R₁、R₄、R₇、R₁₁、R₁₃、R₁₆、R₂₁、R₂₆、R₂₈、R₃₆、R₃₈、R₅₃、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Cysteine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_CYSThe sequence of (SEQ ID NO:574),

wherein R is a ribonucleotide residue and common to Cys is:

R₀is absent

R₁₄、R₃₉、R₅₇Independently is a or absent;

R₄₁a, C or absent;

R₁₀、R₁₅、R₂₇、R₃₃、R₆₂independently A, C, G or absent;

R₃、R₄、R₅、R₆、R₁₂、R₁₃、R₁₆、R₂₄、R₂₆、R₂₉、R₃₀、R₃₁、R₃₂、R₃₄、R₄₂、R₄₄、R₄₅、R₄₆、R₄₈、R₄₉、R₅₈、R₆₃、R₆₄、R₆₆、R₆₇、R₆₈、R₆₉、R₇₀independently N or absent;

R₆₅a, C, U or absent;

R₉、R₂₅、R₃₇、R₄₀、R₅₂、R₅₆independently A, G or absent;

R₇、R₂₀、R₅₁independently A, G, U or absent;

R₁₈、R₃₈、R₅₅independently is C or absent;

R₂c, G or absent;

R₂₁、R₂₈、R₄₃、R₅₀independently C, G, U or absent;

R₁₁、R₂₂、R₂₃、R₃₅、R₃₆、R₅₉、R₆₀、R₆₁、R₇₁、R₇₂independently C, U or absent;

R₁、R₁₉independently G or absent;

R₁₇g, U or absent;

R₈、R₅₃、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_CYSThe sequence of (SEQ ID NO:575),

wherein R is a ribonucleotide residue and common to Cys is:

R₀、R₁₈、R₂₃absent;

R₁₄、R₂₄、R₂₆、R₂₉、R₃₉、R₄₁、R₄₅、R₅₇independently is a or absent;

R₄₄a, C or absent;

R₂₇、R₆₂independently A, C, G or absent;

R₁₆a, C, G, U or absent;

R₃₀、R₇₀independently A, C, U or absent;

R₅、R₇、R₉、R₂₅、R₃₄、R₃₇、R₄₀、R₄₆、R₅₂、R₅₆、R₅₈、R₆₆independently A, G or absent;

R₂₀、R₅₁independently A, G, U or absent;

R₃₅、R₃₈、R₄₃、R₅₅、R₆₉independently is C or absent;

R₂、R₄、R₁₅independently C, G or absent;

R₁₃c, G, U or absent;

R₆、R₁₁、R₂₈、R₃₆、R₄₈、R₄₉、R₅₀、R₆₀、R₆₁、R₆₇、R₆₈、R₇₁、R₇₂independently C, U or absent;

R₁、R₃、R₁₀、R₁₉、R₃₃、R₆₃independently G or absent;

R₈、R₁₇、R₂₁、R₆₄independently G, U or absent;

R₁₂、R₂₂、R₃₁、R₃₂、R₄₂、R₅₃、R₅₄、R₆₅independently is U or absent;

R_59＝u, or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_CYSThe sequence of (SEQ ID NO:576),

wherein R is a ribonucleotide residue and common to Cys is:

R₀、R₁₈、R₂₃is absent

R₁₄、R₂₄、R₂₆、R₂₉、R₃₄、R₃₉、R₄₁、R₄₅、R₅₇、R₅₈Independently is a or absent;

R₄₄、R₇₀independently A, C or absent;

R₆₂a, C, G or absent;

R₁₆n or absent;

R₅、R₇、R₉、R₂₀、R₄₀、R₄₆、R₅₁、R₅₂、R₅₆、R₆₆independently A, G or absent;

R₂₈、R₃₅、R₃₈、R₄₃、R₅₅、R₆₇、R₆₉independently is C or absent;

R₄、R₁₅independently C, G or absent;

R₆、R₁₁、R₁₃、R₃₀、R₄₈、R₄₉、R₅₀、R₆₀、R₆₁、R₆₈、R₇₁、R₇₂independently C, U or absent;

R₁、R₂、R₃、R₁₀、R₁₉、R₂₅、R₂₇、R₃₃、R₃₇、R₆₃independently G or absent;

R₈、R₂₁、R₆₄independently G, U or absent;

R₁₂、R₁₇、R₂₂、R₃₁、R₃₂、R₃₆、R₄₂、R₅₃、R₅₄、R₅₉、R₆₅independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Glutamine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_GLNThe sequence of (SEQ ID NO:577),

wherein R is a ribonucleotide residue and is shared for Gln:

R₀、R₁₈absent;

R₁₄、R₂₄、R₅₇independently is a or absent;

R₉、R₂₆、R₂₇、R₃₃、R₅₆independently A, C, G or absent;

R₂、R₄、R₅、R₆、R₁₂、R₁₃、R₁₆、R₂₁、R₂₂、R₂₅、R₂₉、R₃₀、R₃₁、R₃₂、R₃₄、R₄₁、R₄₂、R₄₄、R₄₅、R₄₆、R₄₈、R₄₉、R₅₀、R₅₈、R₆₂、R₆₃、R₆₆、R₆₇、R₆₈、R₆₉、R₇₀independently N or absent;

R₁₇、R₂₃、R₄₃、R₆₅、R₇₁independently A, C, U or absent;

R₁₅、R₄₀、R₅₁、R₅₂independently A, G or absent;

R₁、R₇、R₇₂independently A, G, U or absent;

R₃、R₁₁、R₃₇、R₆₀、R₆₄independently C, G, U or absent;

R₂₈、R₃₅、R₅₅、R₅₉、R₆₁independently C, U or absent;

R₁₀、R₁₉、R₂₀independently G or absent;

R₃₉g, U or absent;

R₈、R₃₆、R₃₈、R₅₃、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_GLNThe sequence of (SEQ ID NO:578),

wherein R is a ribonucleotide residue and is shared for Gln:

R₀、R₁₈、R₂₃is absent

R₁₄、R₂₄、R₅₇Independently is a or absent;

R₁₇、R₇₁independently A, C or absent;

R₂₅、R₂₆、R₃₃、R₄₄、R₄₆、R₅₆、R₆₉independently A, C, G or absent;

R₄、R₅、R₁₂、R₂₂、R₂₉、R₃₀、R₄₈、R₄₉、R₆₃、R₆₇、R₆₈independently N or absent;

R₃₁、R₄₃、R₆₂、R₆₅、R₇₀independently A, C, U or absent;

R₁₅、R₂₇、R₃₄、R₄₀、R₄₁、R₅₁、R₅₂independently A, G or absent;

R₂、R₇、R₂₁、R₄₅、R₅₀、R₅₈、R₆₆、R₇₂independently A, G, U or absent;

R₃、R₁₃、R₃₂、R₃₇、R₄₂、R₆₀、R₆₄independently C, G, U or absent;

R₆、R₁₁、R₂₈、R₃₅、R₅₅、R₅₉、R₆₁independently C, U or absent;

R₉、R₁₀、R₁₉、R₂₀independently G or absent;

R₁、R₁₆、R₃₉independently G, U or absent;

R₈、R₃₆、R₃₈、R₅₃、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_GLNThe sequence of (SEQ ID NO:579),

wherein R is a ribonucleotide residue and is shared for Gln:

R₀、R₁₈、R₂₃is absent

R₁₄、R₂₄、R₄₁、R₅₇Independently is a or absent;

R₁₇、R₇₁independently A, C or absent;

R₅、R₂₅、R₂₆、R₄₆、R₅₆、R₆₉independently A, C, G or absent;

R₄、R₂₂、R₂₉、R₃₀、R₄₈、R₄₉、R₆₃、R₆₈independently N or absent;

R₄₃、R₆₂、R₆₅、R₇₀independently A, C, U or absent;

R₁₅、R₂₇、R₃₃、R₃₄、R₄₀、R₅₁、R₅₂independently A, G or absent;

R₂、R₇、R₁₂、R₄₅、R₅₀、R₅₈、R₆₆independently A, G, U or absent;

R₃₁a, U or absent;

R₃₂、R₄₄、R₆₀independently C, G or absent;

R₃、R₁₃、R₃₇、R₄₂、R₆₄、R₆₇independently C, G, U or absent;

R₆、R₁₁、R₂₈、R₃₅、R₅₅、R₅₉、R₆₁independently C, U or absent;

R₉、R₁₀、R₁₉、R₂₀independently G or absent;

R₁、R₂₁、R₃₉、R₇₂independently G, U or absent;

R₈、R₁₆、R₃₆、R₃₈、R₅₃、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Glutamic acid TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_GLUThe sequence of (SEQ ID NO:580),

wherein R is a ribonucleotide residue and is common to Glu:

R₀absent;

R₃₄、R₄₃、R₆₈、R₆₉independently A, C, G or absent;

R₁、R₂、R₅、R₆、R₉、R₁₂、R₁₆、R₂₀、R₂₁、R₂₆、R₂₇、R₂₉、R₃₀、R₃₁、R₃₂、R₃₃、R₄₁、R₄₄、R₄₅、R₄₆、R₄₈、R₅₀、R₅₁、R₅₈、R₆₃、R₆₄、R₆₅、R₆₆、R₇₀、R₇₁independently N or absent;

R₁₃、R₁₇、R₂₃、R₆₁independently A, C, U or absent;

R₁₀、R₁₄、R₂₄、R₄₀、R₅₂、R₅₆independently A, G or absent;

R₇、R₁₅、R₂₅、R₆₇、R₇₂independently A, G, U or absent;

R₁₁、R₅₇independently A, U or absent;

R₃₉c, G or absent;

R₃、R₄、R₂₂、R₄₂、R₄₉、R₅₅、R₆₂independently C, G, U or absent;

R₁₈、R₂₈、R₃₅、R₃₇、R₅₃、R₅₉、R₆₀independently C, U or absent;

R₁₉g or absent;

R₈、R₃₆、R₃₈、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_GLUThe sequence of (SEQ ID NO:581),

wherein R is a ribonucleotide residue and is common to Glu:

R₀、R₁₈、R₂₃is absent

R₁₇、R₄₀Independently is a or absent;

R₂₆、R₂₇、R₃₄、R₄₃、R₆₈、R₆₉、R₇₁independently A, C, G or absent;

R₁、R₂、R₅、R₁₂、R₂₁、R₃₁、R₃₃、R₄₁、R₄₅、R₄₈、R₅₁、R₅₈、R₆₆、R₇₀independently N or absent;

R₄₄、R₆₁independently A, C, U or absent;

R₉、R₁₄、R₂₄、R₂₅、R₅₂、R₅₆、R₆₃independently A, G or absent;

R₇、R₁₅、R₄₆、R₅₀、R₆₇、R₇₂independently A, G, U or absent;

R₂₉、R₅₇independently A, U or absent;

R₆₀c or absent;

R₃₉c, G or absent;

R₃、R₆、R₂₀、R₃₀、R₃₂、R₄₂、R₅₅、R₆₂、R₆₅independently C, G, U or absent;

R₄、R₈、R₁₆、R₂₈、R₃₅、R₃₇、R₄₉、R₅₃、R₅₉independently C, U or absent;

R₁₀、R₁₉independently G or absent;

R₂₂、R₆₄independently G, U or absent;

R₁₁、R₁₃、R₃₆、R₃₈、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_GLUThe sequence of (SEQ ID NO:582),

wherein R is a ribonucleotide residue and is common to Glu:

R₀、R₁₇、R₁₈、R₂₃is absent

R₁₄、R₂₇、R₄₀、R₇₁Independently is a or absent;

R₄₄a, C or absent;

R₄₃a, C, G or absent;

R₁、R₃₁、R₃₃、R₄₅、R₅₁、R₆₆independently N or absent;

R₂₁、R₄₁independently A, C, U or absent;

R₇、R₂₄、R₂₅、R₅₀、R₅₂、R₅₆、R₆₃、R₆₈、R₇₀independently A, G or absent;

R₅、R₄₆independently A, G, U or absent;

R₂₉、R₅₇、R₆₇、R₇₂independently A, U or absent;

R₂、R₃₉、R₆₀independently is C or absent;

R₃、R₁₂、R₂₀、R₂₆、R₃₄、R₆₉independently C, G or absent;

R₆、R₃₀、R₄₂、R₄₈、R₆₅independently C, G, Uor is absent;

R₄、R₁₆、R₂₈、R₃₅、R₃₇、R₄₉、R₅₃、R₅₅、R₅₈、R₆₁、R₆₂independently C, U or absent;

R₉、R₁₀、R₁₉、R₆₄independently G or absent;

R₁₅、R₂₂、R₃₂independently G, U or absent;

R₈、R₁₁、R₁₃、R₃₆、R₃₈、R₅₄、R₅₉independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Glycine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_GLYThe sequence of (SEQ ID NO:583),

wherein R is a ribonucleotide residue and shared for Gly is:

R₀absent;

R₂₄a or absent;

R₃、R₉、R₄₀、R₅₀、R₅₁independently A, C, G or absent;

R₄、R₅、R₆、R₇、R₁₂、R₁₆、R₂₁、R₂₂、R₂₆、R₂₉、R₃₀、R₃₁、R₃₂、R₃₃、R₃₄、R₄₁、R₄₂、R₄₃、R₄₄、R₄₅、R₄₆、R₄₈、R₄₉、R₅₈、R₆₃、R₆₄、R₆₅、R₆₆、R₆₇、R₆₈independently N or absent;

R₅₉a, C, U or absent;

R₁、R₁₀、R₁₄、R₁₅、R₂₇、R₅₆independently A, G or absent;

R₂₀、R₂₅independently A, G, U or absent;

R₅₇、R₇₂independently A, U or absent;

R₃₈、R₃₉、R₆₀independently is C or absent;

R₅₂c, G or absent;

R₂、R₁₉、R₃₇、R₅₄、R₅₅、R₆₁、R₆₂、R₆₉、R₇₀independently C, G, U or absent;

R₁₁、R₁₃、R₁₇、R₂₈、R₃₅、R₃₆、R₇₁independently C, U or absent;

R₈、R₁₈、R₂₃、R₅₃independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_GLYThe sequence of (SEQ ID NO:584),

wherein R is a ribonucleotide residue and shared for Gly is:

R_0、R_18、R₂₃is absent

R₂₄、R₂₇、R₄₀、R₇₂Independently is a or absent;

R₂₆a, C or absent;

R₃、R₇、R₆₈independently A, C, G or absent;

R₅、R₃₀、R₄₁、R₄₂、R₄₄、R₄₉、R₆₇independently A, C, G, U or absent;

R₃₁、R₃₂、R₃₄independently A, C, U or absent;

R₉、R₁₀、R₁₄、R₁₅、R₃₃、R₅₀、R₅₆independently A, G or absent;

R₁₂、R₁₆、R₂₂、R₂₅、R₂₉、R₄₆independently A, G, U or absent;

R₅₇a, U or absent;

R₁₇、R₃₈、R₃₉、R₆₀、R₆₁、R₇₁independently is C or absent;

R₆、R₅₂、R₆₄、R₆₆independently C, G or absent;

R₂、R₄、R₃₇、R₄₈、R₅₅、R₆₅independently C, G, U or absent;

R₁₃、R₃₅、R₄₃、R₆₂、R₆₉independently C, U or absent;

R₁、R₁₉、R₂₀、R₅₁、R₇₀independently G or absent;

R₂₁、R₄₅、R₆₃independently G, U or absent;

R₈、R₁₁、R₂₈、R₃₆、R₅₃、R₅₄、R₅₈、R₅₉independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_GLYThe sequence of (SEQ ID NO:585),

wherein R is a ribonucleotide residue and shared for Gly is:

R₀、R_18、R₂₃is absent

R₂₄、R₂₇、R₄₀、R₇₂Independently is a or absent;

R₂₆a, C or absent;

R₃、R₇、R₄₉、R₆₈independently A, C, G or absent;

R₅、R₃₀、R₄₁、R₄₄、R₆₇independently N or absent;

R₃₁、R₃₂、R₃₄independently A, C, U or absent;

R₉、R₁₀、R₁₄、R₁₅、R₃₃、R₅₀、R₅₆independently A, G or absent;

R₁₂、R₂₅、R₂₉、R₄₂、R₄₆independently A, G, U or absent;

R₁₆、R₅₇independently A, U or absent;

R₁₇、R₃₈、R₃₉、R₆₀、R₆₁、R₇₁independently is C or absent;

R₆、R₅₂、R₆₄、R₆₆independently C, G or absent;

R₃₇、R₄₈、R₆₅independently C, G, U or absent;

R₂、R₄、R₁₃、R₃₅、R₄₃、R₅₅、R₆₂、R₆₉independently C, U or absent;

R₁、R₁₉、R₂₀、R₅₁、R₇₀independently G or absent;

R₂₁、R₂₂、R₄₅、R₆₃independently G, U or absent;

R₈、R₁₁、R₂₈、R₃₆、R₅₃、R₅₄、R₅₈、R₅₉independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Histidine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_HISThe sequence of (SEQ ID NO:586),

wherein R is a ribonucleotide residue and is common to His:

R₂₃absent;

R₁₄、R₂₄、R₅₇independently is a or absent;

R₇₂a, C or absent;

R₉、R₂₇、R₄₃、R₄₈、R₆₉independently A, C, G or absent;

R₃、R₄、R₅、R₆、R₁₂、R₂₅、R₂₆、R₂₉、R₃₀、R₃₁、R₃₄、R₄₂、R₄₅、R₄₆、R₄₉、R₅₀、R₅₈、R₆₂、R₆₃、R₆₆、R₆₇、R₆₈independently N or absent;

R₁₃、R₂₁、R₄₁、R₄₄、R₆₅independently A, C, U or absent;

R₄₀、R₅₁、R₅₆、R₇₀independently A, G or absent;

R₇、R₃₂independently A, G, U or absent;

R₅₅、R₆₀independently is C or absent;

R₁₁、R₁₆、R₃₃、R₆₄independently C, G, U or absent;

R₂、R₁₇、R₂₂、R₂₈、R₃₅、R₅₃、R₅₉、R₆₁、R₇₁independently C, U or absent;

R₁、R₁₀、R₁₅、R₁₉、R₂₀、R₃₇、R₃₉、R₅₂independently G or absent;

R₀g, U or absent;

R₈、R₁₈、R₃₆、R₃₈、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_HISThe sequence of (SEQ ID NO:587),

wherein R is a ribonucleotide residue and is common to His:

R₀、R₁₇、R₁₈、R₂₃absent;

R₇、R₁₂、R₁₄、R₂₄、R₂₇、R₄₅、R₅₇、R₅₈、R₆₃、R₆₇、R₇₂independently is a or absent;

R₃a, C, U or absent;

R₄、R₄₃、R₅₆、R₇₀independently A, G or absent;

R₄₉a, U or absent;

R₂、R₂₈、R₃₀、R₄₁、R₄₂、R₄₄、R₄₈、R₅₅、R₆₀、R₆₆、R₇₁independently is C or absent;

R₂₅c, G or absent;

R₉c, G, U or absent;

R₈、R₁₃、R₂₆、R₃₃、R₃₅、R₅₀、R₅₃、R₆₁、R₆₈independently C, U or absent;

R₁、R₆、R₁₀、R₁₅、R₁₉、R₂₀、R₃₂、R₃₄、R₃₇、R₃₉、R₄₀、R₄₆、R₅₁、R₅₂、R₆₅、R₆₄、R₆₉independently G or absent;

R₁₆g, U or absent;

R₅、R₁₁、R₂₁、R₂₂、R₂₉、R₃₁、R₃₆、R₃₈、R₅₄、R₅₉、R₆₅independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_HISThe sequence of (SEQ ID NO:588),

wherein R is a ribonucleotide residue and is common to His:

R₀、R₁₇、R₁₈、R₂₃is absent

R₇、R₁₂、R₁₄、R₂₄、R₂₇、R₄₅、R₅₇、R₅₈、R₆₃、R₆₇、R₇₂Independently is a or absent;

R₃a, C or absent;

R₄、R₄₃、R₅₆、R₇₀independently A, G or absent;

R₄₉a, U or absent;

R₂、R₂₈、R₃₀、R₄₁、R₄₂、R₄₄、R₄₈、R₅₅、R₆₀、R₆₆、R₇₁independently is C or absent;

R₈、R₉、R₂₆、R₃₃、R₃₅、R₅₀、R₆₁、R₆₈independently C, U or absent;

R₁、R₆、R₁₀、R₁₅、R₁₉、R₂₀、R₂₅、R₃₂、R₃₄、R₃₇、R₃₉、R₄₀、R₄₆、R₅₁、R₅₂、R₆₂、R₆₄、R₆₉independently G or absent;

R₅、R₁₁、R₁₃、R₁₆、R₂₁、R₂₂、R₂₉、R₃₁、R₃₆、R₃₈、R₅₃、R₅₄、R₅₉、R₆₅independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Isoleucine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_ILEThe sequence of (SEQ ID NO:589),

wherein R is a ribonucleotide residue and is common to Ile:

R₂₃absent;

R₃₈、R₄₁、R₅₇、R₇₂independently is a or absent;

R₁、R₂₆independently A, C, G or absent;

R₀、R₃、R₄、R₆、R₁₆、R₃₁、R₃₂、R₃₄、R₃₇、R₄₂、R₄₃、R₄₄、R₄₅、R₄₆、R₄₈、R₄₉、R₅₀、R₅₈、R₅₉、R₆₂、R₆₃、R₆₄、R₆₆、R₆₇、R₆₈、R₆₉independently N or absent;

R₂₂、R₆₁、R₆₅independently A, C, U or absent;

R₉、R₁₄、R₁₅、R₂₄、R₂₇、R₄₀independently A, G or absent;

R₇、R₂₅、R₂₉、R₅₁、R₅₆independently A, G, U or absent;

R₁₈、R₅₄independently A, U or absent;

R₆₀c or absent;

R₂、R₅₂、R₇₀independently C, G or absent;

R₅、R₁₂、R₂₁、R₃₀、R₃₃、R₇₁independently C, G, U or absent;

R₁₁、R₁₃、R₁₇、R₂₈、R₃₅、R₅₃、R₅₅independently C, U or absent;

R₁₀、R₁₉、R₂₀independently G or absent;

R₈、R₃₆、R₃₉independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_ILEThe sequence of (SEQ ID NO:590),

wherein R is a ribonucleotide residue and is common to Ile:

R₀、R₁₈、R₂₃is absent

R₂₄、R₃₈、R₄₀、R₄₁、R₅₇、R₇₂Independently is a or absent;

R₂₆、R₆₅independently A, C or absent;

R₅₈、R₅₉、R₆₇independently N or absent;

R₂₂a, C, U or absent;

R₆、R₉、R₁₄、R₁₅、R₂₉、R₃₄、R₄₃、R₄₆、R₄₈、R₅₀、R₅₁、R₆₃、R₆₉independently A, G or absent;

R₃₇、R₅₆independently A, G, U or absent;

R₅₄a, U or absent;

R₂₈、R₃₅、R₆₀、R₆₂、R₇₁independently is C or absent;

R₂、R₅₂、R₇₀independently C, G or absent;

R₅c, G, U or absent;

R₃、R₄、R₁₁、R₁₃、R₁₇、R₂₁、R₃₀、R₄₂、R₄₄、R₄₅、R₄₉、R₅₃、R₅₅、R₆₁、R₆₄、R₆₆independently C, U or absent;

R₁、R₁₀、R₁₉、R₂₀、R₂₅、R₂₇、R₃₁、R₆₈independently G or absent;

R₇、R₁₂、R₃₂independently G, U or absent;

R₈、R₁₆、R₃₃、R₃₆、R₃₉independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_ILEThe sequence of (SEQ ID NO:591),

wherein R is a ribonucleotide residue and is common to Ile:

R₀、R₁₈、R₂₃is absent

R₁₄、R₂₄、R₃₈、R₄₀、R₄₁、R₅₇、R₇₂Independently is a or absent;

R₂₆、R₆₅independently A, C or absent;

R₂₂、R₅₉independently A, C, U or absent;

R₆、R₉、R₁₅、R₃₄、R₄₃、R₄₆、R₅₁、R₅₆、R₆₃、R₆₉independently A, G or absent;

R₃₇a, G, U or absent;

R₁₃、R₂₈、R₃₅、R₄₄、R₅₅、R₆₀、R₆₂、R₇₁independently is C or absent;

R₂、R₅、R₇₀independently C, G or absent;

R₅₈、R₆₇independently C, G, U or absent;

R₃、R₄、R₁₁、R₁₇、R₂₁、R₃₀、R₄₂、R₄₅、R₄₉、R₅₃、R₆₁、R₆₄、R₆₆independently C, U or absent;

R₁、R₁₀、R₁₉、R₂₀、R₂₅、R₂₇、R₂₉、R₃₁、R₃₂、R₄₈、R₅₀、R₅₂、R₆₈independently G or absent;

R₇、R₁₂independently G, U or absent;

R₈、R₁₆、R₃₃、R₃₆、R₃₉、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Methionine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_METThe sequence of (SEQ ID NO:592),

wherein R is a ribonucleotide residue and is common to Met:

R₀、R₂₃absent;

R₁₄、R₃₈、R₄₀、R₅₇independently is a or absent;

R₆₀a, C or absent;

R₃₃、R₄₈、R₇₀independently A, C, G or absent;

R₁、R₃、R₄、R₅、R₆、R₁₁、R₁₂、R₁₆、R₁₇、R₂₁、R₂₂、R₂₆、R₂₇、R₂₉、R₃₀、R₃₁、R₃₂、R₄₂、R₄₄、R₄₅、R₄₆、R₄₉、R₅₀、R₅₈、R₆₂、R₆₃、R₆₆、R₆₇、R₆₈、R₆₉、R₇₁independently N or absent;

R₁₈、R₃₅、R₄₁、R₅₉、R₆₅independently A, C, U or absent;

R₉、R₁₅、R₅₁independently A, G or absent;

R₇、R₂₄、R₂₅、R₃₄、R₅₃、R₅₆independently A, G, U or absent;

R₇₂a, U or absent;

R₃₇c or absent;

R₁₀、R₅₅independently C, G or absent;

R₂、R₁₃、R₂₈、R₄₃、R₆₄independently C, G, U or absent;

R₃₆、R₆₁independently C, U or absent;

R₁₉、R₂₀、R₅₂independently G or absent;

R₈、R₃₉、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_METThe sequence of (SEQ ID NO:593),

wherein R is a ribonucleotide residue and is common to Met:

R₀、R₁₈、R₂₂、R₂₃is absent

R₁₄、R₂₄、R₃₈、R₄₀、R₄₁、R₅₇、R₇₂Independently is a or absent;

R₅₉、R₆₀、R₆₂、R₆₅independently A, C or absent;

R₆、R₄₅、R₆₇independently A, C, G or absent;

R₄n or absent;

R₂₁、R₄₂independently A, C, U or absent;

R₁、R₉、R₂₇、R₂₉、R₃₂、R₄₆、R₅₁independently A, G or absent;

R₁₇、R₄₉、R₅₃、R₅₆、R₅₈independently A, G, U or absent;

R₆₃a, U or absent;

R₃、R₁₃、R₃₇independently is C or absent;

R₄₈、R₅₅、R₆₄、R₇₀independently C, G or absent;

R₂、R₅、R₆₆、R₆₈independently C, G, U or absent;

R₁₁、R₁₆、R₂₆、R₂₈、R₃₀、R₃₁、R₃₅、R₃₆、R₄₃、R₄₄、R₆₁、R₇₁independently C, U or absent;

R₁₀、R₁₂、R₁₅、R₁₉、R₂₀、R₂₅、R₃₃、R₅₂、R₆₉independently G or absent;

R₇、R₃₄、R₅₀independently G, U or absent;

R₈、R₃₉、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_METThe sequence of (SEQ ID NO:594),

wherein R is a ribonucleotide residue and is common to Met:

R₀、R₁₈、R₂₂、R₂₃is absent

R₁₄、R₂₄、R₃₈、R₄₀、R₄₁、R₅₇、R₇₂Independently is a or absent;

R₅₉、R₆₂、R₆₅independently A, C or absent;

R₆、R₆₇independently A, C, G or absent;

R₄、R₂₁independently A, C, U or absent;

R₁、R₉、R₂₇、R₂₉、R₃₂、R₄₅、R₄₆、R₅₁independently A, G or absent;

R₁₇、R₅₆、R₅₈independently A, G, U or absent;

R₄₉、R₅₃、R₆₃independently A, U or absent;

R₃、R₁₃、R₂₆、R₃₇、R₄₃、R₆₀independently is C or absent;

R₂、R₄₈、R₅₅、R₆₄、R₇₀independently C, G or absent;

R₅、R₆₆independently C, G, U or absent;

R₁₁、R₁₆、R₂₈、R₃₀、R₃₁、R₃₅、R₃₆、R₄₂、R₄₄、R₆₁、R₇₁independently C, U or absent;

R₁₀、R₁₂、R₁₅、R₁₉、R₂₀、R₂₅、R₃₃、R₅₂、R₆₉independently G or absent;

R₇、R₃₄、R₅₀、R₆₈independently G, U or absent;

R₈、R₃₉、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Leucine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_LEUThe sequence of (SEQ ID NO:595),

wherein R is a ribonucleotide residue and is shared with Leu:

R₀absent;

R₃₈、R₅₇independently is a or absent;

R₆₀a, C or absent;

R₁、R₁₃、R₂₇、R₄₈、R₅₁、R₅₆independently A, C, G or absent;

R₂、R₃、R₄、R₅、R₆、R₇、R₉、R₁₀、R₁₁、R₁₂、R₁₆、R₂₃、R₂₆、R₂₈、R₂₉、R₃₀、R₃₁、R₃₂、R₃₃、R₃₄、R₃₇、R₄₁、R₄₂、R₄₃、R₄₄、R₄₅、R₄₆、R₄₉、R₅₀、R₅₈、R₆₂、R₆₃、R₆₅、R₆₆、R₆₇、R₆₈、R₆₉、R₇₀independently N or absent;

R₁₇、R₁₈、R₂₁、R₂₂、R₂₅、R₃₅、R₅₅independently A, C, U or absent;

R₁₄、R₁₅、R₃₉、R₇₂independently A, G or absent;

R₂₄、R₄₀independently A, G, U or absent;

R₅₂、R₆₁、R₆₄、R₇₁independently C, G, U or absent;

R₃₆、R₅₃、R₅₉independently C, U or absent;

R₁₉g or absent;

R₂₀g, U or absent;

R₈、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_LEUThe sequence of (SEQ ID NO:596),

wherein R is a ribonucleotide residue and is shared with Leu:

R₀is absent

R₃₈、R₅₇、R₇₂Independently is a or absent;

R₆₀a, C or absent;

R₄、R₅、R₄₈、R₅₀、R₅₆、R₆₉independently A, C, G or absent;

R₆、R₃₃、R₄₁、R₄₃、R₄₆、R₄₉、R₅₈、R₆₃、R₆₆、R₇₀independently N or absent;

R₁₁、R₁₂、R₁₇、R₂₁、R₂₂、R₂₈、R₃₁、R₃₇、R₄₄、R₅₅independently A, C, U or absent;

R₁、R₉、R₁₄、R₁₅、R₂₄、R₂₇、R₃₄、R₃₉independently A, G or absent;

R₇、R₂₉、R₃₂、R₄₀、R₄₅independently A, G, U or absent;

R₂₅a, U or absent;

R₁₃c, G or absent;

R₂、R₃、R₁₆、R₂₆、R₃₀、R₅₂、R₆₂、R₆₄、R₆₅、R₆₇、R₆₈independently C, G, U or absent;

R₁₈、R₃₅、R₄₂、R₅₃、R₅₉、R₆₁、R₇₁independently C, U or absent;

R₁₉、R₅₁independently G or absent;

R₁₀、R₂₀independently G, U or absent;

R₈、R₂₃、R₃₆、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_LEUThe sequence of (SEQ ID NO:597),

wherein R is a ribonucleotide residue and is shared with Leu:

R₀is absent

R₃₈、R₅₇、R₇₂Independently is a or absent;

R₆₀a, C or absent;

R₄、R₅、R₄₈、R₅₀、R₅₆、R₅₈、R₆₉independently A, C, G or absent;

R₆、R₃₃、R₄₃、R₄₆、R₄₉、R₆₃、R₆₆、R₇₀independently N or absent;

R₁₁、R₁₂、R₁₇、R₂₁、R₂₂、R₂₈、R₃₁、R₃₇、R₄₁、R_44、R₅₅independently A, C, U or absent;

R₁、R₉、R₁₄、R₁₅、R₂₄、R₂₇、R₃₄、R₃₉independently A, G or absent;

R₇、R₂₉、R₃₂、R₄₀、R₄₅independently A, G, U or absent;

R₂₅a, U or absent;

R₁₃c, G or absent;

R₂、R₃、R₁₆、R₃₀、R₅₂、R₆₂、R₆₄、R₆₇、R₆₈independently C, G, U or absent;

R₁₈、R₃₅、R₄₂、R₅₃、R₅₉、R₆₁、R₆₅、R₇₁independently C, U or absent;

R₁₉、R₅₁independently G or absent;

R₁₀、R₂₀、R₂₆independently G, U or absent;

R₈、R₂₃、R₃₆、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Lysine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_LYSSequence of (SEQ ID NO: 598))，

Wherein R is a ribonucleotide residue and is common to Lys is:

R₀is absent

R₁₄A or absent;

R₄₀、R₄₁independently A, C or absent;

R₃₄、R₄₃、R₅₁independently A, C, G or absent;

R₁、R₂、R₃、R₄、R₅、R₆、R₇、R₁₁、R₁₂、R₁₆、R₂₁、R₂₆、R₃₀、R₃₁、R₃₂、R₄₄、R₄₅、R₄₆、R₄₈、R₄₉、R₅₀、R₅₈、R₆₂、R₆₃、R₆₅、R₆₆、R₆₇、R₆₈、R₆₉、R₇₀independently N or absent;

R₁₃、R₁₇、R₅₉、R₇₁independently A, C, U or absent;

R₉、R₁₅、R₁₉、R₂₀、R₂₅、R₂₇、R₅₂、R₅₆independently A, G or absent;

R₂₄、R₂₉、R₇₂independently A, G, U or absent;

R₁₈、R₅₇independently A, U or absent;

R₁₀、R₃₃independently C, G or absent;

R₄₂、R₆₁、R₆₄independently C, G, U or absent;

R₂₈、R₃₅、R₃₆、R₃₇、R₅₃、R₅₅、R₆₀independently C, U or absent;

R₈、R₂₂、R₂₃、R₃₈、R₃₉、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_LYSThe sequence of (SEQ ID NO:599),

wherein R is a ribonucleotide residue and is common to Lys is:

R₀、R₁₈、R₂₃is absent

R₁₄A or absent;

R₄₀、R₄₁、R₄₃independently A, C or absent;

R₃、R₇independently A, C, G or absent;

R₁、R₆、R₁₁、R₃₁、R₄₅、R₄₈、R₄₉、R₆₃、R₆₅、R₆₆、R₆₈independently N or absent;

R₂、R₁₂、R₁₃、R₁₇、R₄₄、R₆₇、R₇₁independently A, C, U or absent;

R₉、R₁₅、R₁₉、R₂₀、R₂₅、R₂₇、R₃₄、R₅₀、R₅₂、R₅₆、R₇₀、R₇₂independently A, G or absent;

R₅、R₂₄、R₂₆、R₂₉、R₃₂、R₄₆、R₆₉independently A, G, U or absent;

R₅₇a, U or absent;

R₁₀、R₆₁independently C, G or absent;

R₄、R₁₆、R₂₁、R₃₀、R₅₈、R₆₄independently C, G, U or absent;

R₂₈、R₃₅、R₃₆、R₃₇、R₄₂、R₅₃、R₅₅、R₅₉、R₆₀、R₆₂independently C, U or absent;

R₃₃、R₅₁independently G or absent;

R₈g, U or absent;

R₂₂、R₃₈、R₃₉、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_LYSThe sequence of (SEQ ID NO:600),

wherein R is a ribonucleotide residue and is common to Lys is:

R₀、R₁₈、R₂₃is absent

R₉、R₁₄、R₃₄、R₄₁Independently is a or absent;

R₄₀a, C or absent;

R₁、R₃、R₇、R₃₁independently A, C, G or absent;

R₄₈、R₆₅、R₆₈independently N or absent;

R₂、R₁₃、R₁₇、R₄₄、R₆₃、R₆₆independently A, C, U or absent;

R₅、R₁₅、R₁₉、R₂₀、R₂₅、R₂₇、R₂₉、R₅₀、R₅₂、R₅₆、R₇₀、R₇₂independently A, G or absent;

R₆、R₂₄、R₃₂、R₄₉independently A, G, U or absent;

R₁₂、R₂₆、R₄₆、R₅₇independently A, U or absent;

R₁₁、R₂₈、R₃₅、R₄₃independently is C or absent;

R₁₀、R₄₅、R₆₁independently C, G or absent;

R₄、R₂₁、R₆₄independently C, G, U or absent;

R₃₇、R₅₃、R₅₅、R₅₉、R₆₀、R₆₂、R₆₇、R₇₁independently C, U or absent;

R₃₃、R₅₁independently G or absent;

R₈、R₃₀、R₅₈、R₆₉independently G, U or absent;

R₁₆、R₂₂、R₃₆、R₃₈、R₃₉、R₄₂、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Phenylalanine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_PHEThe sequence of (SEQ ID NO:601),

wherein R is a ribonucleotide residue and is common to Phe:

R₀、R₂₃is absent

R₉、R₁₄、R₃₈、R₃₉、R₅₇、R₇₂Independently is a or absent;

R₇₁a, C or absent;

R₄₁、R₇₀independently A, C, G or absent;

R₄、R₅、R₆、R₃₀、R₆₁、R₃₂、R₃₄、R₄₂、R₄₄、R₄₅、R₄₆、R₄₈、R₄₉、R₅₈、R₆₂、R₆₃、R₆₆、R₆₇、R₆₈、R₆₉independently N or absent;

R₁₆、R₆₁、R₆₅independently A, C, U or absent;

R₁₅、R₂₆、R₂₇、R₂₉、R₄₀、R₅₆independently A, G or absent;

R₇、R₅₁independently A, G, U or absent;

R₂₂、R₂₄independently A, U or absent;

R₅₅、R₆₀independently is C or absent;

R₂、R₃、R₂₁、R₃₃、R₄₃、R₅₀、R₆₄independently C, G, U or absent;

R₁₁、R₁₂、R₁₃、R₁₇、R₂₈、R₃₅、R₃₆、R₅₉independently C, U or absent;

R₁₀、R₁₉、R₂₀、R₂₅、R₃₇、R₅₂independently G or absent;

R₁g, U or absent;

R₈、R₁₈、R₅₃、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_PHEThe sequence of (SEQ ID NO:602),

wherein R is a ribonucleotide residue and is common to Phe:

R₀、R₁₈、R₂₃is absent

R₁₄、R₂₄、R₃₈、R₃₉、R₅₇、R₇₂Independently is a or absent;

R₄₆、R₇₁independently A, C or absent;

R₄、R₇₀independently A, C, G or absent;

R₄₅a, C, U or absent;

R₆、R₇、R₁₅、R₂₆、R₂₇、R₃₂、R₃₄、R₄₀、R₄₁、R₅₆、R₆₉independently A, G or absent;

R₂₉a, G, U or absent;

R₅、R₉、R₆₇independently A, U or absent;

R₃₅、R₄₉、R₅₅、R₆₀independently is C or absent;

R₂₁、R₄₃、R₆₂independently C, G or absent;

R₂、R₃₃、R₆₈independently C, G, U or absent;

R₃、R₁₁、R₁₂、R₁₃、R₂₈、R₃₀、R₃₆、R₄₂、R₄₄、R₄₈、R₅₈、R₅₉、R₆₁、R₆₆independently C, U or absent;

R₁₀、R₁₉、R₂₀、R₂₅、R₃₇、R₅₁、R₅₂、R₆₃、R₆₄independently G or absent;

R₁、R₃₁、R₅₀independently G, U or absent;

R₈、R₁₆、R₁₇、R₂₂、R₅₃、R₅₄、R₆₅independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_PHEThe sequence of (SEQ ID NO:603),

wherein R is a ribonucleotide residue and is common to Phe:

R₀、R₁₈、R₂₂、R₂₃is absent

R₅、R₇、R₁₄、R₂₄、R₂₆、R₃₂、R₃₄、R₃₈、R₃₉、R₄₁、R₅₇、R₇₂Independently is a or absent;

R₄₆a, C or absent;

R₇₀a, C, G or absent;

R₄、R₆、R₁₅、R₅₆、R₆₉independently A, G or absent;

R₉、R₄₅independently A, U or absent;

R₂、R₁₁、R₁₃、R₃₅、R₄₃、R₄₉、R₅₅、R₆₀、R₆₈、R₇₁independently is C or absent;

R₃₃c, G or absent;

R₃、R₂₈、R₃₆、R₄₈、R₅₈、R₅₉、R₆₁independently C, U or absent;

R₁、R₁₀、R₁₉、R₂₀、R₂₁、R₂₅、R₂₇、R₂₉、R₃₇、R₄₀、R₅₁、R₅₂、R₆₂、R₆₃、R₆₄independently G or absent;

R₈、R₁₂、R₁₆、R₁₇、R₃₀、R₃₁、R₄₂、R₄₄、R₅₀、R₅₃、R₅₄、R₆₅、R₆₆、R₆₇independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Proline TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_PROThe sequence of (SEQ ID NO:604),

wherein R is a ribonucleotide residue and is common to Pro:

R₀is absent

R₁₄、R₅₇Independently is a or absent;

R₇₀、R₇₂independently A, C or absent;

R₉、R₂₆、R₂₇independently A, C, G or absent;

R₄、R₅、R₆、R₁₆、R₂₁、R₂₉、R₃₀、R₃₁、R₃₂、R₃₃、R₃₄、R₃₇、R₄₁、R₄₂、R₄₃、R₄₄、R₄₅、R₄₆、R₄₈、R₄₉、R₅₀、R₅₈、R₆₁、R₆₂、R₆₃、R₆₄、R₆₆、R₆₇、R₆₈independently N or absent;

R₃₅、R₆₅independently A, C, U or absent;

R₂₄、R₄₀、R₅₆independently A, G or absent;

R₇、R₂₅、R₅₁independently A, G, U or absent;

R₅₅、R₆₀independently is C or absent;

R₁、R₃、R₇₁independently C, G or absent;

R₁₁、R₁₂、R₂₀、R₆₉independently C, G, U or absent;

R₁₃、R₁₇、R₁₈、R₂₂、R₂₃、R₂₈、R₅₉independently C, U or absent;

R₁₀、R₁₅、R₁₉、R₃₈、R₃₉、R₅₂independently G or absent;

R₂independently G, U or absent;

R₈、R₃₆、R₅₃、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_PROThe sequence of (SEQ ID NO:605),

wherein R is a ribonucleotide residue and is common to Pro:

R₀、R₁₇、R₁₈、R₂₂、R₂₃absent;

R₁₄、R₄₅、R₅₆、R₅₇、R₅₈、R₆₅、R₆₈independently is a or absent;

R₆₁a, C, G or absent;

R₄₃n or absent;

R₃₇a, C, U or absent;

R₂₄、R₂₇、R₃₃、R₄₀、R₄₄、R₆₃independently A, G or absent;

R₃、R₁₂、R₃₀、R₃₂、R₄₈、R₅₅、R₆₀、R₇₀、R₇₁、R₇₂independently is C or absent;

R₅、R₃₄、R₄₂、R₆₆independently C, G or absent;

R₂₀c, G, U or absent;

R₃₅、R₄₁、R₄₉、R₆₂independently C, U or absent;

R₁、R₂、R₆、R₉、R₁₀、R₁₅、R₁₉、R₂₆、R₃₈、R₃₉、R₄₆、R₅₀、R₅₁、R₅₂、R₆₄、R₆₇、R₆₉independently G or absent;

R₁₁、R₁₆independently G, U or absent;

R₄、R₇、R₈、R₁₃、R₂₁、R₂₅、R₂₈、R₂₉、R₃₁、R₃₆、R₅₃、R₅₄、R₅₉independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_PROThe sequence of (SEQ ID NO:606),

wherein R is a ribonucleotide residue and is common to Pro:

R₀、R₁₇、R₁₈、R₂₂、R₂₃is absent

R₁₄、R₄₅、R₅₆、R₅₇、R₅₈、R₆₅、R₆₈Independently is a or absent;

R₃₇a, C, U or absent;

R₂₄、R₂₇、R₄₀independently A, G or absent;

R₃、R₅、R₁₂、R₃₀、R₃₂、R₄₈、R₄₉、R₅₅、R₆₀、R₆₁、R₆₂、R₆₆、R₇₀、R₇₁、R₇₂independently is C or absent;

R₃₄、R₄₂independently C, G or absent;

R₄₃c, G, U or absent;

R₄₁c, U or absent;

R₁、R₂、R₆、R₉、R₁₀、R₁₅、R₁₉、R₂₀、R₂₆、R₃₃、R₃₈、R₃₉、R₄₄、R₄₆、R₅₀、R₅₁、R₅₂、R₆₃、R₆₄、R₆₇、R₆₉independently G or absent;

R₁₆g, U or absent;

R₄、R₇、R₈、R₁₁、R₁₃、R₂₁、R₂₅、R₂₈、R₂₉、R₃₁、R₃₅、R₃₆、R₅₃、R₅₄、R₅₉independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Serine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_SERThe sequence of (SEQ ID NO:607),

wherein R is a ribonucleotide residue and common to Ser is:

R₀absent;

R₁₄、R₂₄、R₅₇independently is a or absent;

R₄₁a, C or absent;

R₂、R₃、R₄、R₅、R₆、R₇、R₉、R₁₀、R₁₁、R₁₂、R₁₃、R₁₆、R₂₁、R₂₅、R₂₆、R₂₇、R₂₈、R₃₀、R₃₁、R₃₂、R₃₃、R₃₄、R₃₇、R₄₂、R₄₃、R₄₄、R₄₅、R₄₆、R₄₈、R₄₉、R₅₀、R₆₂、R₆₃、R₆₄、R₆₅、R₆₆、R₆₇、R₆₈、R₆₉、R₇₀independently N or absent;

R₁₈a, C, U or absent;

R₁₅、R₄₀、R₅₁、R₅₆independently isA. G or is absent;

R₁、R₂₉、R₅₈、R₇₂independently A, G, U or absent;

R₃₉a, U or absent;

R₆₀c or absent;

R₃₈c, G or absent;

R₁₇、R₂₂、R₂₃、R₇₁independently C, G, U or absent;

R₈、R₃₅、R₃₆、R₅₅、R₅₉、R₆₁independently C, U or absent;

R₁₉、R₂₀independently G or absent;

R₅₂g, U or absent;

R₅₃、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_SERThe sequence of (SEQ ID NO:608),

wherein R is a ribonucleotide residue and common to Ser is:

R₀、R₂₃is absent

R₁₄、R₂₄、R₄₁、R₅₇Independently is a or absent;

R₄₄a, C or absent;

R₂₅、R₄₅、R₄₈independently A, C, G or absent;

R₂、R₃、R₄、R₅、R₃₇、R₅₀、R₆₂、R₆₆、R₆₇、R₆₉、R₇₀independently N or absent;

R₁₂、R₂₈、R₆₅independently A, C, U or absent;

R₉、R₁₅、R₂₉、R₃₄、R₄₀、R₅₆、R₆₃independently A, G or absent;

R₇、R₂₆、R₃₀、R₃₃、R₄₆、R₅₈、R₇₂independently A, G, U or absent;

R₃₉a, U or absent;

R₁₁、R₃₅、R₆₀、R₆₁independently is C or absent;

R₁₃、R₃₈independently C, G or absent;

R₆、R₁₇、R₃₁、R₄₃、R₆₄、R₆₈independently C, G, U or absent;

R₃₆、R₄₂、R₄₉、R₅₅、R₅₉、R₇₁independently C, U or absent;

R₁₀、R₁₉、R₂₀、R₂₇、R₅₁independently G or absent;

R₁、R₁₆、R₃₂、R₅₂independently G, U or absent;

R₈、R₁₈、R₂₁、R₂₂、R₅₃、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_SERThe sequence of (SEQ ID NO:609),

wherein R is a ribonucleotide residue and common to Ser is:

R₀、R₂₃is absent

R₁₄、R₂₄、R₄₁、R₅₇、R₅₈Independently is a or absent;

R₄₄a, C or absent;

R₂₅、R₄₈independently A, C, G or absent;

R₂、R₃、R₅、R₃₇、R₆₆、R₆₇、R₆₉、R₇₀independently N or absent;

R₁₂、R₂₈、R₆₂independently A, C, U or absent;

R₇、R₉、R₁₅、R₂₉、R₃₃、R₃₄、R₄₀、R₄₅、R₅₆、R₆₃independently A, G or absent;

R₄、R₂₆、R₄₆、R₅₀independently A, G, U or absent;

R₃₀、R₃₉independently A, U or absent;

R₁₁、R₁₇、R₃₅、R₆₀、R₆₁independently is C or absent;

R₁₃、R₃₈independently C, G or absent;

R₆、R₆₄independently C, G, U or absent;

R₃₁、R₄₂、R₄₃、R₄₉、R₅₅、R₅₉、R₆₅、R₆₈、R₇₁independently C, U or absent;

R₁₀、R₁₉、R₂₀、R₂₇、R₅₁、R₅₂independently G or absent;

R₁、R₁₆、R₃₂、R₇₂independently G, U or absent;

R₈、R₁₈、R₂₁、R₂₂、R₃₆、R₅₃、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Threonine TREM consensus sequences

In embodiments, a TREM disclosed herein comprises formula I_THRThe sequence of (SEQ ID NO:610),

wherein R is a ribonucleotide residue and is common to Thr:

R₀、R₂₃is absent

R₁₄、R₄₁、R₅₇Independently is a or absent;

R₅₆、R₇₀independently A, C, G or absent;

R₄、R₅、R₆、R₇、R₁₂、R₁₆、R₂₆、R₃₀、R₃₁、R₃₂、R₃₄、R₃₇、R₄₂、R₄₄、R₄₅、R₄₆、R₄₈、R₄₉、R₅₀、R₅₈、R₆₂、R₆₃、R₆₄、R₆₅、R₆₆、R₆₇、R₆₈、R₇₂independently N or absent;

R₁₃、R₁₇、R₂₁、R₃₅、R₆₁independently A, C, U or absent;

R₁、R₉、R₂₄、R₂₇、R₂₉、R₆₉independently A, G or absent;

R₁₅、R₂₅、R₅₁independently A, G, U or absent;

R₄₀、R₅₃independently A, U or absent;

R₃₃、R₄₃independently C, G or absent;

R₂、R₃、R₅₉independently C, G, U or absent;

R₁₁、R₁₈、R₂₂、R₂₈、R₃₆、R₅₄、R₅₅、R₆₀、R₇₁independently C, U or absent;

R₁₀、R₂₀、R₃₈、R₅₂independently G or absent;

R₁₉g, U or absent;

R₈、R₃₉independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_THRThe sequence of (SEQ ID NO:611),

wherein R is a ribonucleotide residue and is common to Thr:

R₀、R₁₈、R₂₃is absent

R₁₄、R₄₁、R₅₇Independently is a or absent;

R₉、R₄₂、R₄₄、R₄8、R₅₆、R₇₀independently A, C, G or absent;

R₄、R₆、R₁₂、R₂₆、R₄₉、R₅₈、R₆₃、R₆₄、R₆₆、R₆₈independently N or absent;

R₁₃、R₂₁、R₃₁、R₃₇、R₆₂independently A, C, U or absent;

R₁、R₁₅、R₂₄、R₂₇、R₂₉、R₄₆、R₅₁、R₆₉independently A, G or absent;

R₇、R₂₅、R₄₅、R₅₀、R₆₇independently A, G, U or absent;

R₄₀、R₅₃independently A, U or absent;

R₃₅c or absent;

R₃₃、R₄₃independently C, G or absent;

R₂、R₃、R₅、R₁₆、R₃₂、R₃₄、R₅₉、R₆₅、R₇₂independently C, G, U or absent;

R₁₁、R₁₇、R₂₂、R₂₈、R₃₀、R₃₆、R₅₅、R₆₀、R₆₁、R₇₁independently C, U or absent;

R₁₀、R₁₉、R₂₀、R₃₈、R₅₂independently G or absent;

R₈、R₃₉、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_THRThe sequence of (SEQ ID NO:612),

wherein R is a ribonucleotide residue and is common to Thr:

R₀、R₁₈、R₂₃is absent

R₁₄、R₄₀、R₄₁、R₅₇Independently is a or absent;

R₄₄a, C or absent;

R₉、R₄₂、R₄₈、R₅₆independently A, C, G or absent;

R₄、R₆、R₁₂、R₂₆、R₅₈、R₆₄、R₆₆、R₆₈independently N or absent;

R₁₃、R₂₁、R₃₁、R₃₇、R₄₉、R₆₂independently A, C, U or absent;

R₁、R₁₅、R₂₄、R₂₇、R₂₉、R₄₆、R₅₁、R₆₉independently A, G or absent;

R₇、R₂₅、R₄₅、R₅₀、R₆₃、R₆₇independently A, G, U or absent;

R₅₃a, U or absent;

R₃₅c or absent;

R₂、R₃₃、R₄₃、R₇₀independently C, G or absent;

R₅、R₁₆、R₃₄、R₅₉、R₆₅independently C, G, U or absent;

R₃、R₁₁、R₂₂、R₂₈、R₃₀、R₃₆、R₅₅、R₆₀、R₆₁、R₇₁independently C, U or absent;

R₁₀、R₁₉、R₂₀、R₃₈、R₅₂independently G or absent;

R₃₂g, U or absent;

R₈、R₁₇、R₃₉、R₅₄、R₇₂independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Tryptophan TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_TRPThe sequence of (SEQ ID NO:613),

wherein R is a ribonucleotide residue and is shared for Trp:

R₀absent;

R₂₄、R₃₉、R₄₁、R₅₇independently is a or absent;

R₂、R₃、R₂₆、R₂₇、R₄₀、R₄₈independently A, C, G or absent;

R₄、R₅、R₆、R₂₉、R₃₀、R₃₁、R₃₂、R₃₄、R₄₂、R₄₄、R₄₅、R₄₆、R₄₉、R₅₁、R₅₈、R₆₃、R₆₆、R₆₇、R₆₈independently N or absent;

R₁₃、R₁₄、R₁₆、R₁₈、R₂₁、R₆₁、R₆₅、R₇₁independently A, C, U or absent;

R₁、R₉、R₁₀、R₁₅、R₃₃、R₅₀、R₅₆independently A, G or absent;

R₇、R₂₅、R₇₂independently A, G, U or absent;

R₃₇、R₃₈、R₅₅、R₆₀independently is C or absent;

R₁₂、R₃₅、R₄₃、R₆₄、R₆₉、R₇₀independently C, G, U or absent;

R₁₁、R₁₇、R₂₂、R₂₈、R₅₉、R₆₂independently C, U or absent;

R₁₉、R₂₀、R₅₂independently G or absent;

R₈、R₂₃、R₃₆、R₅₃、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_TRPThe sequence of (SEQ ID NO:614),

wherein R is a ribonucleotide residue and is shared for Trp:

R₀、R₁₈、R₂₂、R₂₃is absent

R₁₄、R₂₄、R₃₉、R₄₁、R₅₇、R₇₂Independently is a or absent;

R₃、R₄、R₁₃、R₆₁、R₇₁independently A, C or absent;

R₆、R₄₄independently A, C, G or absent;

R₂₁a, C, U or absent;

R₂、R₇、R₁₅、R₂₅、R₃₃、R₃₄、R₄₅、R₅₆、R₆₃independently A, G or absent;

R₅₈a, G, U or absent;

R₄₆a, U or absent;

R₃₇、R₃₈、R₅₅、R₆₀、R₆₂independently is C or absent;

R₁₂、R₂₆、R₂₇、R₃₅、R₄₀、R₄₈、R₆₇independently C, G or absent;

R₃₂、R₄₃、R₆₈independently C, G, U or absent;

R₁₁、R₁₆、R₂₈、R₃₁、R₄₉、R₅₉、R₆₅、R₇₀independently C, U or absent;

R₁、R₉、R₁₀、R₁₉、R₂₀、R₅₀、R₅₂、R₆₉independently G or absent;

R₅、R₈、R₂₉、R₃₀、R₄₂、R₅₁、R₆₄、R₆₆independently G, U or absent;

R₁₇、R₃₆、R₅₃、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_TRPThe sequence of (SEQ ID NO:615),

wherein R is a ribonucleotide residue and is shared for Trp:

R₀、R₁₈、R₂₂、R₂₃is absent

R₁₄、R₂₄、R₃₉、R₄₁、R₅₇、R₇₂Independently is a or absent;

R₃、R₄、R₁₃、R₆₁、R₇₁independently A, C or absent;

R₆、R₄₄independently A, C, G or absent;

R₂₁a, C, U or absent;

R₂、R₇、R₁₅、R₂₅、R₃₃、R₃₄、R₄₅、R₅₆、R₆₃independently A, G or absent;

R₅₈a, G, U or absent;

R₄₆a, U or absent;

R₃₇、R₃₈、R₅₅、R₆₀、R₆₂independently is C or absent;

R₁₂、R₂₆、R₂₇、R₃₅、R₄₀、R₄₈、R₆₇independently C, G or absent;

R₃₂、R₄₃、R₆₈independently C, G, U or absent;

R₁₁、R₁₆、R₂₈、R₃₁、R₄₉、R₅₉、R₆₅、R₇₀independently C, U or absent;

R₁、R₉、R₁₀、R₁₉、R₂₀、R₅₀、R₅₂、R₆₉independently G or absent;

R₅、R₈、R₂₉、R₃₀、R₄₂、R₅₁、R₆₄、R₆₆independently G, U or absent;

R₁₇、R₃₆、R₅₃、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Tyrosine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_TYRThe sequence of (SEQ ID NO:616),

wherein R is a ribonucleotide residue and shared for Tyr is:

R₀is absent

R₁₄、R₃₉、R₅₇Independently is a or absent;

R₄₁、R₄₈、R₅₁、R₇₁independently A, C, G or absent;

R₃、R₄、R₅、R₆、R₉、R₁₀、R₁₂、R₁₃、R₁₆、R₂₅、R₂₆、R₃₀、R₃₁、R₃₂、R₄₂、R₄₄、R₄₅、R₄₆、R₄₉、R₅₀、R₅₈、R₆₂、R₆₃、R₆₆、R₆₇、R₆₈、R₆₉、R₇₀independently N or absent;

R₂₂、R₆₅independently A, CU or absent;

R₁₅、R₂₄、R₂₇、R₃₃、R₃₇、R₄₀、R₅₆independently A, G or absent;

R₇、R₂₉、R₃₄、R₇₂independently A, G, U or absent;

R₂₃、R₅₃independently A, U or absent;

R₃₅、R₆₀independently is C or absent;

R₂₀c, G or absent;

R₁、R₂、R₂₈、R₆₁、R₆₄independently C, G, U or absent;

R₁₁、R₁₇、R₂₁、R₄₃、R₅₅independently C, U or absent;

R₁₉、R₅₂independently G or absent;

R₈、R₁₈、R₃₆、R₃₈、R₅₄、R₅₉independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_TYRThe sequence of (SEQ ID NO:617),

wherein R is a ribonucleotide residue and shared for Tyr is:

R₀、R₁₈、R₂₃is absent

R₇、R₉、R₁₄、R₂₄、R₂₆、R₃₄、R₃₉、R₅₇Independently is a or absent；

R₄₄、R₆₉Independently A, C or absent;

R₇₁a, C, G or absent;

R₆₈n or absent;

R₅₈a, C, U or absent;

R₃₃、R₃₇、R₄₁、R₅₆、R₆₂、R₆₃independently A, G or absent;

R₆、R₂₉、R₇₂independently A, G, U or absent;

R₃₁、R₄₅、R₅₃independently A, U or absent;

R₁₃、R₃₅、R₄₉、R₆₀independently is C or absent;

R₂₀、R₆₈、R₆₄、R₆₇、R₇₀independently C, G or absent;

R₁、R₂、R₅、R₁₆、R₆₆independently C, G, U or absent;

R₁₁、R₂₁、R₂₈、R₄₃、R₅₅、R₆₁independently C, U or absent;

R₁₀、R₁₅、R₁₉、R₂₅、R₂₇、R₄₀、R₅₁、R₅₂independently G or absent;

R₃、R₄、R₃₀、R₃₂、R₄₂、R₄₆independently G, U or absent;

R₈、R₁₂、R₁₇、R₂₂、R₃₆、R₃₈、R₅₀、R₅₄、R₅₉、R₆₅independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_TYRThe sequence of (SEQ ID NO:618),

wherein R is a ribonucleotide residue and shared for Tyr is:

R₀、R₁₈、R₂₃is absent

R₇、R₉、R₁₄、R₂₄、R₂₆、R₃₄、R₃₉、R₅₇、R₇₂Independently is a or absent;

R₄₄、R₆₉independently A, C or absent;

R₇₁a, C, G or absent;

R₃₇、R₄₁、R₅₆、R₆₂、R₆₃independently A, G or absent;

R₆、R₂₉、R₆₈independently A, G, U or absent;

R₃₁、R₄₅、R₅₈independently A, U or absent;

R₁₃、R₂₈、R₃₅、R₄₉、R₆₀、R₆₁independently is C or absent;

R₅、R₄₈、R₆₄、R₃₇、R₇₀independently C, G or absent;

R₁、R₂independently C, G, U or absent;

R₁₁、R₁₆、R₂₁、R₄₃、R₅₅、R₆₆independently C, U or absent;

R₁₀、R₁₅、R₁₉、R₂₀、R₂₅、R₂₇、R₃₃、R₄₀、R₅₁、R₅₂independently G or absent;

R₃、R₄、R₃₀、R₃₂、R₄₂、R₄₆independently G, U or absent;

R₈、R₁₂、R₁₇、R₂₂、R₃₆、R₃₈、R₅₀、R₅₃、R₅₄、R₅₉、R₆₅independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Valine TREM consensus sequence

In embodiments, a TREM disclosed herein comprises formula I_VALThe sequence of (SEQ ID NO:619),

wherein R is a ribonucleotide residue and shared for Val is:

R₀、R₂₃absent;

R₂₄、R₃₈、R₅₇independently is a or absent;

R₉、R₇₂independently A, C, G or absent;

R₂、R₄、R₅、R₆、R₇、R₁₂、R₁₅、R₁₆、R₂₁、R₂₅、R₂₆、R₂₉、R₃₁、R₃₂、R₃₃、R₃₄、R₃₇、R₄₁、R₄₂、R₄₃、R₄₄、R₄₅、R₄₆、R₄₈、R₄₉、R₅₀、R₅₈、R₆₁、R₆₂、R₆₃、R₆₄、R₆₅、R₆₆、R₆₇、R₆₈、R₆₉、R₇₀independently N or absent;

R₁₇、R₃₅、R₅₉independently A, C, U or absent;

R₁₀、R₁₄、R₂₇、R₄₀、R₅₂、R₅₆independently A, G or absent;

R₁、R₃、R₅₁、R₅₃independently A, G, U or absent;

R₃₉c or absent;

R₁₃、R₃₀、R₅₅independently C, G, U or absent;

R₁₁、R₂₂、R₂₈、R₆₀、R₇₁independently C, U or absent;

R₁₉g or absent;

R₂₀g, U or absent;

R₈、R₁₈、R₃₆、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula II_VALThe sequence of (SEQ ID NO:620),

wherein R is a ribonucleotide residue and shared for Val is:

R₀、R₁₈、R₂₃absent;

R₂₄、R₃₈、R₅₇independently is a or absent;

R₆₄、R₇₀、R₇₂either aloneEither immediately A, C, G or absent;

R₁₅、R₁₆、R₂₆、R₂₉、R₃₁、R₃₂、R₄₃、R₄₄、R₄₅、R₄₉、R₅₀、R₅₈、R₆₂、R₆₅independently N or absent;

R₆、R₁₇、R₃₄、R₃₇、R₄₁、R₅₉independently A, C, U or absent;

R₉、R₁₀、R₁₄、R₂₇、R₄₀、R₄₆、R₅₁、R₅₂、R₅₆independently A, G or absent;

R₇、R₁₂、R₂₅、R₃₃、R₅₃、R₆₃、R₆₆、R₆₈independently A, G, U or absent;

R₆₉a, U or absent;

R₃₉c or absent;

R₅、R₆₇independently C, G or absent;

R₂、R₄、R₁₃、R₄₈、R₅₅、R₆₁independently C, G, U or absent;

R₁₁、R₂₂、R₂₈、R₃₀、R₃₅、R₆₀、R₇₁independently C, U or absent;

R₁₉g or absent;

R₁、R₃、R₂₀、R₄₂independently G, U or absent;

R₈、R₂₁、R₃₆、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

In embodiments, a TREM disclosed herein comprises formula III_VALThe sequence of (SEQ ID NO:621),

wherein R is a ribonucleotide residue and shared for Val is:

R₀、R₁₈、R₂₃is absent

R₂₄、R₃₈、R₄₀、R₅₇、R₇₂Independently is a or absent;

R₂₉、R₆₄、R₇₀independently A, C, G or absent;

R₄₉、R₅₀、R₆₂independently N or absent;

R₁₆、R₂₆、R₃₁、R₃₂、R₃₇、R₄₁、R₄₃、R₅₉、R₆₅independently A, C, U or absent;

R₉、R₁₄、R₂₇、R₄₆、R₅₂、R₅₆、R₆₆independently A, G or absent;

R₇、R₁₂、R₂₅、R₃₃、R₄₄、R₄₅、R₅₃、R₅₈、R₆₃、R₆₈independently A, G, U or absent;

R₆₉a, U or absent;

R₃₉c or absent;

R₅、R₆₇independently C, G or absent;

R₂、R₄、R₁₃、R₁₅、R₄₈、R₅₅independently C, G, U or absent;

R₆、R₁₁、R₂₂、R₂₈、R₃₀、R₃₄、R₃₅、R₆₀、R₆₁、R₇₁independently C, U or absent;

R₁₀、R₁₉、R₅₁independently G or absent;

R₁、R₃、R₂₀、R₄₂independently G, U or absent;

R₈、R₁₇、R₂₁、R₃₆、R₅₄independently is U or absent;

[R₄₇]_xn or absent;

Provided that TREM has one or both of the following characteristics: no more than 15% of the residues are N; or no more than 20 residues are absent.

Variable region consensus sequences

In embodiments, TREM disclosed herein is at position R₄₇And includes a variable region. In embodiments, the variable region is 1-271 ribonucleotides (e.g., 1-250, 1-22) in length5. 1-200, 1-175, 1-150, 1-125, 1-100, 1-75, 1-50, 1-40, 1-30, 1-29, 1-28, 1-27, 1-26, 1-25, 1-24, 1-23, 1-22, 1-21, 1-20, 1-19, 1-18, 1-17, 1-16, 1-15, 1-14, 1-13, 1-12, 1-11, 1-10, 10-271, 20-271, 30-271, 40-271, 50-271, 60-271, 70-271, 80-271, 100-271, 125-271, 150-271, 175-271, 200-271, 225-271, 1. 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 50, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200, 225, 250, or 271 ribonucleotides). In embodiments, the variable region comprises any one, all, or a combination of adenine, cytosine, guanine, or uracil.

In embodiments, the variable region comprises a ribonucleic acid (RNA) sequence encoded by a deoxyribonucleic acid (DNA) sequence disclosed in Table 3 (e.g., any one of SEQ ID NOS: 452-561 disclosed in Table 3).

Table 3: exemplary variable region sequences.

Method for preparing TREM

Methods for designing and constructing expression vectors and modifying host cells for production of a target (e.g., a TREM or enzyme disclosed herein) use techniques known in the art. For example, cells are genetically modified to express an exogenous TREM using cultured mammalian cells (e.g., cultured human cells), insect cells, yeast, bacteria, or other cells under the control of an appropriate promoter. Generally, recombinant methods can be used. See generally Pharmaceutical Biotechnology, fundametals and Applications [ Pharmaceutical Biotechnology: foundation and applications ], Springer [ sporling press ] (2013); green and Sambrook (eds.), Molecular Cloning: A Laboratory Manual [ Molecular Cloning-A Laboratory Manual ] (fourth edition), Cold Spring Harbor Laboratory Press [ Cold Spring Harbor Laboratory Press ] (2012). For example, mammalian expression vectors can contain non-transcribed elements, such as origins of replication, suitable promoters and enhancers, and other 5 'or 3' flanking non-transcribed sequences. DNA sequences derived from the SV40 viral genome, such as the SV40 origin, early promoter, enhancer, splicing and polyadenylation sites may be used to provide additional genetic elements required for expression of the heterologous DNA sequence.

Methods of making a TREM or TREM composition disclosed herein include the use of a host cell, e.g., a modified host cell, that expresses TREM.

Culturing the modified host cell under conditions that allow expression of TREM. In embodiments, the culture conditions can be adjusted to increase expression of TREM. The method of producing a TREM further comprises purifying the expressed TREM from the host cell culture to produce a TREM composition. In embodiments, the TREM is a TREM fragment, e.g., a tRNA fragment encoded by a deoxyribonucleic acid sequence disclosed in table 1. For example, a TREM includes less than the entire sequence of a tRNA from the same species as the subject being treated, e.g., less than the entire sequence of a tRNA having the same anticodon, or both. In embodiments, production of TREM fragments, e.g., from full-length TREM or longer fragments, can be catalyzed by an enzyme, e.g., an enzyme having nuclease activity (e.g., endonuclease activity or ribonuclease activity), such as rnase a, dicer, angiogenin, rnase P, RNA enzyme Z, Rny1, or PrrC.

In embodiments, a method of making a TREM described herein comprises contacting (e.g., transducing or transfecting) a host cell (e.g., as described herein, e.g., a modified host cell) with an exogenous nucleic acid (e.g., DNA or RNA) encoding a TREM described herein under conditions sufficient for expression of the TREM. In embodiments, the exogenous nucleic acid comprises RNA (or DNA encoding RNA) comprising a ribonucleic acid (RNA) sequence of RNA encoded by a DNA sequence disclosed in table 1. In embodiments, the exogenous nucleic acid comprises an RNA sequence (or DNA encoding an RNA sequence) that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an RNA sequence encoded by a DNA sequence provided in table 1. In embodiments, the exogenous nucleic acid comprises an RNA sequence (or DNA encoding an RNA sequence) comprising at least 30 consecutive nucleotides of a ribonucleic acid (RNA) sequence encoded by a deoxyribonucleic acid (DNA) sequence disclosed in table 1. In embodiments, the exogenous nucleic acid comprises an RNA sequence (or DNA encoding an RNA sequence) comprising at least 30 contiguous nucleotides of an RNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an RNA sequence encoded by a DNA sequence provided in table 1.

In an example, the host cell is transduced with a virus (e.g., lentivirus, adenovirus, or retrovirus) that expresses TREM, as described in example 8.

The expressed TREM can be purified from the host cell or host cell culture to produce a TREM composition, e.g., as described herein. Purification of TREM can be performed by affinity purification, e.g., as described in MACS isolation protocols for particular tRNA molecules, or other methods known in the art. In the examples, TREM was purified by the method described in example 7.

In embodiments, a method of making a TREM, e.g., a TREM composition, includes contacting a TREM with an agent (e.g., a capture reagent comprising a nucleic acid sequence complementary to a TREM). A single capture reagent or multiple capture reagents can be used to prepare a TREM, e.g., a TREM composition. When a single capture reagent is used, the capture reagent can have at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% complementary sequence to TREM. When multiple capture reagents are used, TREM compositions having multiple different TREMs can be prepared. In embodiments, the capture reagent may be coupled to an agent, such as biotin.

In embodiments, the method comprises denaturing the TREM, e.g., prior to hybridization to the capture reagent. In embodiments, the method comprises renaturing TREM after hybridization and/or release from the capture reagent.

In embodiments, a method of preparing a TREM, e.g., a TREM composition, includes contacting the TREM with a reagent (e.g., a separation reagent, e.g., a chromatography reagent). In an embodiment, the chromatography reagent comprises a column chromatography reagent, a planar chromatography reagent, a displacement chromatography reagent, a gas chromatography reagent, a liquid chromatography reagent, an affinity chromatography reagent, an ion exchange chromatography reagent, or a size exclusion agent chromatography reagent.

In embodiments, the TREM prepared by any of the methods described herein can be: (i) a load amino acid, e.g., a homologous amino acid; (ii) (ii) loaded with a non-homologous amino acid (e.g., a misloaded TREM (mTREM); or (iii) unloaded with an amino acid, e.g., an unloaded TREM (uTREM)).

In embodiments, the TREM prepared by any of the methods described herein is an unsupported TREM (utrem). In an embodiment, a method of making a uTREM includes culturing a host cell in a medium having a limiting amount of one or more nutrients (e.g., a media nutrient deficiency).

In embodiments, a loaded TREM, e.g., a TREM loaded with homologous AA or non-homologous AA, can be unloaded, e.g., by dissociating AA, e.g., by incubating the TREM at an elevated temperature.

Exogenous nucleic acids encoding TREM or TREM fragments

In embodiments, the exogenous nucleic acid, e.g., DNA or RNA encoding a TREM, comprises a nucleic acid sequence comprising one or more RNA sequences encoded by a DNA sequence disclosed in table 1 (e.g., any one of SEQ ID NOs 1-451 disclosed in table 1). In embodiments, the exogenous nucleic acid, e.g., a DNA or RNA encoding a TREM, comprises a core sequence that is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence disclosed in table 1 (e.g., any one of SEQ ID NOs 1-451 disclosed in table 1). In one embodiment, the exogenous nucleic acid, e.g., DNA or RNA encoding a TREM, comprises a nucleic acid sequence that is less than 100% identical to an RNA sequence encoded by a DNA sequence disclosed in Table 1 (e.g., any one of SEQ ID NOS: 1-451 disclosed in Table 1).

In embodiments, the exogenous nucleic acid, e.g., DNA or RNA encoding a TREM, comprises a nucleic acid sequence of an RNA sequence encoded by a DNA sequence disclosed in Table 1 (e.g., any one of SEQ ID NOS: 1-451 disclosed in Table 1). In embodiments, the exogenous nucleic acid, e.g., DNA or RNA encoding a TREM, comprises a nucleic acid sequence that is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a plurality of RNA sequences encoded by a DNA sequence disclosed in table 1 (e.g., any of SEQ ID NOs 1-451 disclosed in table 1). In embodiments, the exogenous nucleic acid encoding a TREM comprises an RNA sequence encoded by a DNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence disclosed in table 1 (e.g., any one of SEQ ID NOs 1-451 disclosed in table 1). In embodiments, the exogenous nucleic acid encoding a TREM comprises an RNA sequence encoded by a DNA sequence that is less than 100% identical to a DNA sequence disclosed in table 1 (e.g., any one of SEQ ID NOs 1-451 disclosed in table 1).

In embodiments, the exogenous nucleic acid, e.g., DNA or RNA encoding a TREM, comprises an RNA sequence of one or more TREM fragments, e.g., a fragment of an RNA encoded by a DNA sequence disclosed in table 1, e.g., a fragment of any one of SEQ ID NOs 1-451 as disclosed herein, e.g., in table 1. In embodiments, a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% of the nucleic acid sequence of an RNA encoded by a DNA sequence provided in table 1 (e.g., any one of SEQ ID NOs: 1-451 disclosed in table 1). In embodiments, a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, or 99% of the nucleic acid sequence that is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to an RNA encoded by a DNA sequence provided in table 1. In embodiments, a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% of the nucleic acid sequence encoded by a DNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in table 1 (e.g., any of SEQ ID NOs 1-451 disclosed in table 1).

In embodiments, a TREM fragment comprises at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 consecutive nucleotides of an RNA sequence encoded by a DNA sequence disclosed in table 1 (e.g., any one of SEQ ID NOs 1-451 disclosed in table 1). In embodiments, a TREM fragment comprises at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 consecutive nucleotides of an RNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence disclosed in table 1 (e.g., any one of SEQ ID NOs 1-451 disclosed in table 1). In embodiments, a TREM fragment comprises 26, 27, 28, 29, or 30 consecutive nucleotides of an RNA sequence encoded by a DNA sequence that is at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in table 1 (e.g., any one of SEQ ID NOs 1-451 disclosed in table 1), 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25).

In embodiments, the exogenous nucleic acid comprises DNA that expresses TREM post-transcriptionally.

In embodiments, the exogenous nucleic acid comprises RNA that, upon reverse transcription, produces DNA that can be transcribed to provide a TREM.

In embodiments, the exogenous nucleic acid encoding a TREM comprises: (i) a control region sequence; (ii) a sequence encoding a modified TREM; (iii) a sequence encoding more than one TREM; or (iv) tRNA^MetSequences other than the sequence.

In embodiments, the exogenous nucleic acid encoding a TREM comprises a promoter sequence. In embodiments, the exogenous nucleic acid comprises an RNA polymerase III (Pol III) recognition sequence, e.g., a Pol III binding sequence. In embodiments, the promoter sequence comprises the U6 promoter sequence or a fragment thereof. In embodiments, the nucleic acid sequence comprises a promoter sequence comprising a mutation, e.g., a promoter up-regulation mutation, e.g., a mutation that increases transcription initiation, e.g., a mutation that increases TFIIIB binding. In embodiments, the nucleic acid sequence comprises a promoter sequence that increases Pol III binding and results in increased tRNA production (e.g., TREM production).

Also disclosed herein are plasmids comprising an exogenous nucleic acid encoding a TREM. In embodiments, the plasmid comprises a promoter sequence, e.g., as described herein.

TREM composition

In embodiments, a TREM composition, e.g., a TREM pharmaceutical composition, comprises a pharmaceutically acceptable excipient. Exemplary excipients include those provided in the FDA inactive ingredients database (https:// www.accessdata.fda.gov/scripts/cder/iig/index. cfm).

In embodiments, a TREM composition, e.g., a TREM pharmaceutical composition, comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100, or 150 grams TREM. In embodiments, a TREM composition, e.g., a TREM pharmaceutical composition, comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, or 100 milligrams of TREM.

In embodiments, a TREM composition, e.g., a TREM pharmaceutical composition, is at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or 99% dry weight TREM.

In embodiments, the TREM composition comprises at least 1x 10⁶A TREM molecule of at least 1x 10⁷A TREM molecule of at least1x 10⁸A TREM molecule or at least 1x 10⁹A TREM molecule.

In embodiments, a TREM composition produced by any of the methods of preparation disclosed herein can be loaded with an amino acid using an in vitro loading reaction as disclosed in example 11 or known in the art.

In embodiments, the TREM composition comprises one species or a plurality of species of TREM. In embodiments, the TREM composition comprises a single species of TREM. In embodiments, the TREM composition comprises a first TREM species and a second TREM species. In embodiments, the TREM composition comprises an XTREM species, wherein X ═ 2, 3, 4, 5, 6, 7, 8, 9, or 10.

In embodiments, the TREM is at least 70%, 75%, 80%, 85%, 90%, or 95% identical, or 100% identical, to a sequence encoded by a nucleic acid in table 1.

In embodiments, TREM comprises a consensus sequence provided herein.

The TREM composition can be formulated as a liquid composition, a lyophilized composition, or a frozen composition.

In some embodiments, the TREM composition can be formulated to be suitable for pharmaceutical use, e.g., a pharmaceutical TREM composition. In embodiments, the pharmaceutical TREM composition is substantially free of materials and/or reagents used for isolating and/or purifying TREM, such as the isolating reagents described herein.

In some embodiments, the TREM composition can be formulated with water for injection. In some embodiments, TREM compositions formulated with water for injection are suitable for pharmaceutical use, e.g., comprising a pharmaceutical TREM composition.

TREM purification

TREM compositions, such as TREM pharmaceutical compositions, can be purified from host cells by nucleotide purification techniques. In one embodiment, the TREM composition is purified by affinity purification (e.g., as described in the MACS isolation protocol for a particular tRNA molecule) or by the methods described in examples 1-3 or 7. In one embodiment, the TREM composition is purified by liquid chromatography, e.g., reverse phase ion pair chromatography (IP-RP), ion exchange chromatography (IE), Affinity Chromatography (AC), Size Exclusion Chromatography (SEC), and combinations thereof. See, for example, Baronti et al Analytical and Bioanalytical Chemistry [ Analytical and Bioanalytical Chemistry ] (2018)410: 3239-.

In embodiments, a TREM composition can be purified using a purification method that includes one, two, or all of the following steps, e.g., in the order recited: (i) separating the nucleic acids from the proteins to provide an RNA preparation; (ii) separating RNA of less than 200nt from larger RNA species; and/or (iii) separating TREM from other RNA species by affinity-based, e.g., sequence affinity, separation.

In an embodiment, steps (i) - (iii) are performed in the order recited.

In an embodiment, the purification method comprises step (i). In embodiments, step (i) comprises extracting nucleic acids from proteins in the sample, e.g., as described in example 1. In the examples, the extraction method comprises phenol chloroform extraction,

In an embodiment, the purification method comprises step (ii). In an embodiment, step (ii) is performed on the sample after step (i). In embodiments, step (ii) comprises separating RNA of less than a threshold size, e.g., less than 500nt, 400nt, 300nt, 250nt, or 200nt from larger RNA, e.g., using a miRNeasy kit as described in example 1. In embodiments, step (ii) comprises performing salt precipitation, e.g., LiCl precipitation, to enrich for small RNAs (e.g., remove large RNAs), as described in example 1. In embodiments, separation of RNA of less than a threshold size from larger RNA is performed prior to salt precipitation, e.g., LiCl precipitation, e.g., using miRNeasy kit. In embodiments, step (ii) further comprises performing a desalting or buffer exchange step, for example using a G25 column.

In an embodiment, the purification method comprises step (iii). In embodiments, step (iii) comprises performing affinity-based separation to enrich the TREM. In an embodiment, step (iii) is performed on the sample after step (i) and/or step (ii). In embodiments, affinity-based separations include sequence-based separations, e.g., using probes (e.g., oligonucleotides) comprising sequences that bind TREMs, e.g., as described in example 1. In embodiments, the probe (e.g., oligonucleotide) comprises one or more tags, such as a biotin tag and/or a fluorescent tag.

In embodiments, a TREM purification process comprising steps (i), (ii), and (iii) produces a purified TREM composition. In embodiments, TREM compositions purified according to the methods described herein result in fewer RNA contaminants, e.g., as compared to Trizol RNA extraction purification methods.

TREM quality control and production evaluation

A TREM or TREM composition (e.g., a pharmaceutical TREM composition produced by any of the methods disclosed herein) can be assessed for a characteristic associated with the TREM or TREM preparation, such as purity, host cell protein or DNA content, endotoxin level, sterility, TREM concentration, TREM structure, or functional activity of TREM. Any of the above characteristics can be assessed by providing a value for the characteristic, for example, by assessing or testing TREM, a TREM composition, or an intermediate in the production of a TREM composition. The values may also be compared to standard or reference values. Responsive to the evaluation, the TREM composition may be classified, for example, ready for release, compliance with manufacturing standards for human testing, compliance with ISO standards, compliance with cGMP standards, or compliance with other pharmaceutical standards. In response to the evaluation, the TREM composition can be further processed, e.g., it can be aliquoted, e.g., divided into single or multiple doses, placed in a container (e.g., an end-use vial), packaged, shipped, or placed into commerce. In embodiments, in response to the assessment, one or more characteristics can be adjusted, processed, or reprocessed to optimize the TREM composition. For example, a TREM composition can be conditioned, processed, or reprocessed to (i) increase the purity of the TREM composition; (ii) reducing the amount of HCP in the composition; (iii) reducing the amount of DNA in the composition; (iv) reducing the amount of fragments in the composition; (v) reducing the amount of endotoxin in the composition; (vi) increasing the in vitro translation activity of the composition; (vii) increasing the TREM concentration of the composition; or (viii) inactivate or remove any viral contaminants present in the composition, for example by lowering the pH of the composition or by filtration.

In embodiments, a TREM (e.g., a TREM composition or an intermediate in the production of a TREM composition) has a purity, i.e., by mass, of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.

In embodiments, a TREM (e.g., a TREM composition or an intermediate in the production of a TREM composition) has less than 0.1ng/ml, 1ng/ml, 5ng/ml, 10ng/ml, 15ng/ml, 20ng/ml, 25ng/ml, 30ng/ml, 35ng/ml, 40ng/ml, 50ng/ml, 60ng/ml, 70ng/ml, 80ng/ml, 90ng/ml, 100ng/ml, 200ng/ml, 300ng/ml, 400ng/ml, or 500ng/ml of Host Cell Protein (HCP) contamination.

In an embodiment, a TREM (e.g., a TREM composition or an intermediate in the production of a TREM composition) has less than 0.1ng, 1ng, 5ng, 10ng, 15ng, 20ng, 25ng, 30ng, 35ng, 40ng, 50ng, 60ng, 70ng, 80ng, 90ng, 100ng, 200ng, 300ng, 400ng, or 500ng per milligram (mg) of a TREM composition of Host Cell Protein (HCP) contamination.

In embodiments, a TREM (e.g., a TREM composition or an intermediate in the production of a TREM composition) has a DNA content, e.g., host cell DNA content, of less than 1ng/ml, 5ng/ml, 10ng/ml, 15ng/ml, 20ng/ml, 25ng/ml, 30ng/ml, 35ng/ml, 40ng/ml, 50ng/ml, 60ng/ml, 70ng/ml, 80ng/ml, 90ng/ml, 100ng/ml, 200ng/ml, 300ng/ml, 400ng/ml, or 500 ng/ml.

In embodiments, a TREM (e.g., a TREM composition or an intermediate in the production of a TREM composition) has less than 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25% TREM fragments relative to full-length TREM.

In embodiments, a TREM (e.g., a TREM composition or an intermediate in the production of a TREM composition) has a low level or absence of endotoxin, e.g., a negative result as measured by a Limulus Amebocyte Lysate (LAL) test;

in embodiments, a TREM (e.g., a TREM composition or an intermediate in the production of a TREM composition) has in vitro translation activity as measured by the assay described in example 15.

In embodiments, a TREM (e.g., a TREM composition or an intermediate in the production of a TREM composition) has a TREM concentration of at least 0.1ng/mL, 0.5ng/mL, 1ng/mL, 5ng/mL, 10ng/mL, 50ng/mL, 0.1ug/mL, 0.5ug/mL, 1ug/mL, 2ug/mL, 5ug/mL, 10ug/mL, 20ug/mL, 30ug/mL, 40ug/mL, 50ug/mL, 60ug/mL, 70ug/mL, 80ug/mL, 100ug/mL, 200ug/mL, 300ug/mL, 500ug/mL, 1000ug/mL, 5000ug/mL, 10,000ug/mL, or 100,000 ug/mL.

In embodiments, a TREM (e.g., a TREM composition or an intermediate in the production of a TREM composition) is sterile, e.g., the composition or formulation supports the growth of less than 100 viable microorganisms when tested under sterile conditions, the composition or formulation meets USP <71> standards, and/or the composition or formulation meets USP <85> standards.

In embodiments, a TREM (e.g., a TREM composition or an intermediate in the production of a TREM composition) has an undetectable level of viral contamination, e.g., is free of viral contamination. In embodiments, any viral contaminants present in the composition, such as residual viruses, are inactivated or removed. In embodiments, any viral contaminants, such as residual virus, are inactivated, for example by lowering the pH of the composition. In embodiments, any viral contaminants, such as residual viruses, are removed, such as by filtration or other methods known in the art.

TREM administration

A TREM composition or pharmaceutical composition described herein can be administered to a cell, tissue, or subject, for example, by direct administration to a cell, tissue, and/or organ in vitro, ex vivo, or in vivo. In vivo administration may be via, for example, local, systemic and/or parenteral routes, such as intravenous, subcutaneous, intraperitoneal, intrathecal, intramuscular, ocular, nasal, urogenital, intradermal, dermal, enteral, intravitreal, intracerebral, intrathecal or epidural routes.

Carrier and vehicle

In some embodiments, a TREM or TREM composition described herein is delivered to a cell, e.g., a mammalian cell or a human cell, using a vector. The vector may be, for example, a plasmid or a virus. In some embodiments, the delivery is in vivo, in vitro, ex vivo, or in situ. In some embodiments, the virus is an adeno-associated virus (AAV), lentivirus, adenovirus. In some embodiments, the system or components of the system are delivered to the cell with the virus-like particle or virion. In some embodiments, delivery uses more than one virus, virus-like particle, or virosome.

Carrier

The TREM, TREM composition, or pharmaceutical TREM composition described herein can comprise a carrier, can be formulated with a carrier, or can be delivered in a carrier.

Viral vectors

The carrier can be a viral vector (e.g., a viral vector comprising a sequence encoding a TREM). The viral vector can be administered to a cell or subject (e.g., a human subject or an animal model) to deliver a TREM, a TREM composition, or a pharmaceutical TREM composition. The viral vector may be administered systemically or locally (e.g., by injection).

The viral genome provides a rich source of vectors that can be used to efficiently deliver foreign genes into mammalian cells. It is known in the art that viral genomes can be used as useful vectors for delivery, as polynucleotides contained in such genomes are typically incorporated into the nuclear genome of mammalian cells by universal or specialized transduction. These processes are part of the natural viral replication cycle and do not require the addition of proteins or agents to induce gene integration. Examples of viral vectors include retroviruses (e.g., retroviral vectors of the family retroviral family), adenoviruses (e.g., Ad5, Ad26, Ad34, Ad35, and Ad48), parvoviruses (e.g., adeno-associated viruses), coronaviruses, negative strand RNA viruses (such as orthomyxoviruses (e.g., influenza viruses), rhabdoviruses (e.g., rabies and vesicular stomatitis viruses), paramyxoviruses (e.g., measles and sendai viruses)), positive strand RNA viruses (e.g., picornaviruses and alphaviruses), and double stranded DNA viruses (including adenoviruses, herpesviruses (e.g., herpes simplex virus types 1 and 2, epstein-barr virus, cytomegalovirus, replication-defective herpesviruses), and poxviruses (e.g., vaccinia, Modified Vaccinia (MVA), fowlpox, and canarypox)). Other viruses include, for example, norwalk virus, togavirus, flavivirus, reovirus, papova virus, hepatitis virus, human papilloma virus, human foamy virus, and hepatitis virus. Examples of retroviruses include: avian leukosarcoma, avian type C viruses, mammalian type C, type B viruses, type D viruses, cancer retroviruses, HTLV-BLV groups, lentiviruses, alpha-retroviruses, gamma-retroviruses, foamy viruses (coffee, J.M., Retrovidae: The viruses and replication thereof, Virology [ Virology ] (third edition) Lippincott-Raven, Philadelphia, 1996). Other examples include murine leukemia virus, murine sarcoma virus, mouse mammary tumor virus, bovine leukemia virus, feline sarcoma virus, avian leukemia virus, human T cell leukemia virus, baboon endogenous virus, gibbon ape leukemia virus, Meisengenshui (Mason Pfizer) monkey virus, monkey immunodeficiency virus, monkey sarcoma virus, rous sarcoma virus, and lentiviruses. Other examples of vectors are described, for example, in U.S. Pat. No. 5,801,030, the teachings of which are incorporated herein by reference. In some embodiments, the system or components of the system are delivered to the cell with the virus-like particle or virion.

Cell and vesicle based carriers

TREM, TREM compositions, or pharmaceutical TREM compositions described herein can be administered to cells in vesicles or other membrane-based carriers.

In embodiments, a TREM or TREM composition or pharmaceutical TREM composition described herein is administered in or via a cell, vesicle, or other membrane-based carrier. In one embodiment, the TREM or TREM composition or pharmaceutical TREM composition may be formulated in liposomes or other similar vesicles. Liposomes are spherical vesicular structures consisting of a monolayer or multilamellar lipid bilayer surrounding an inner aqueous compartment and a relatively impermeable outer lipophilic phospholipid bilayer. Liposomes can be anionic, neutral, or cationic. Liposomes are biocompatible, non-toxic, and can deliver hydrophilic and lipophilic Drug molecules, protect their cargo from degradation by plasma enzymes, and transport their cargo across biological membranes and the Blood Brain Barrier (BBB) (for reviews, see, e.g., Spuch and Navarro, Journal of Drug Delivery [ Journal of Drug Delivery ], volume 2011, article ID469679, page 12, 2011.doi: 10.1155/2011/469679).

Vesicles can be made from several different types of lipids; however, phospholipids are most commonly used to generate liposomes as drug carriers. Methods of preparing multilamellar vesicle lipids are known in the art (see, e.g., U.S. patent No. 6,693,086, the teachings of which are incorporated herein by reference for the preparation of multilamellar vesicle lipids). Although vesicle formation is spontaneous when the lipid membrane is mixed with an aqueous solution, vesicle formation can also be accelerated by applying force in the form of shaking using a homogenizer, sonicator or a squeezing device (for review, see, for example, Spuch and Navarro, Journal of Drug Delivery, vol.2011, article ID469679, p.12, 2011.doi: 10.1155/2011/469679). Extruded lipids can be prepared by extrusion through filters of reduced size, as described in Templeton et al, Nature Biotech [ Nature Biotech ],15:647-652,1997, the teachings of which with respect to extruded lipid preparation are incorporated herein by reference.

Lipid nanoparticles are another example of a carrier that provides a biocompatible and biodegradable delivery system for a TREM or TREM composition or a pharmaceutical TREM composition described herein. Nanostructured Lipid Carriers (NLCs) are modified Solid Lipid Nanoparticles (SLNs) that retain the properties of SLNs, improve drug stability and drug loading, and prevent drug leakage. Polymeric Nanoparticles (PNPs) are an important component of drug delivery. These nanoparticles can effectively direct drug delivery to specific targets and improve drug stability and controlled drug release. Lipopolymer Nanoparticles (PLN), a novel carrier that combines liposomes and polymers, can also be used. These nanoparticles have the complementary advantages of PNP and liposomes. PLN consists of a core-shell structure; the polymer core provides a stable structure and the phospholipid shell provides good biocompatibility. Thus, the two components improve the efficiency of drug encapsulation, facilitate surface modification, and prevent leakage of water-soluble drugs. For reviews, see, e.g., Li et al 2017, Nanomaterials [ Nanomaterials ]7,122; doi:10.3390/nano 7060122.

Exosomes may also be used as drug delivery vehicles for TREMs or TREM compositions or pharmaceutical TREM compositions described herein. For a review, see Ha et al, 2016, 7 months, Acta pharmaceutical Sinica B. [ Pharmacology paper ] Vol.6, No. 4, pp 287-296; https:// doi.org/10.1016/j.apsb.2016.02.001.

Ex vivo differentiated red blood cells can also be used as a carrier for a TREM or TREM composition or a pharmaceutical TREM composition described herein. See, for example, WO 2015073587; WO 2017123646; WO 2017123644; WO 2018102740; wO 2016183482; WO 2015153102; WO 2018151829; WO 2018009838; shi et al 2014, Proc Natl Acad Sci USA [ Proc Natl Acad of sciences USA ]111(28) 10131-; us patent 9,644,180; huang et al 2017 Nature Communications [ Nature Communications ]8: 423; shi et al 2014, Proc Natl Acad Sci USA [ Proc. Natl. Acad. Sci. USA ]111(28) 10131-.

The fusion compositions, for example as described in WO 2018208728, can also be used as a carrier to deliver a TREM or TREM composition or a pharmaceutical TREM composition as described herein.

Use of TREM

A TREM composition (e.g., a pharmaceutical TREM composition described herein) can modulate a function of a cell, tissue, or subject. In embodiments, a TREM composition described herein (e.g., a pharmaceutical TREM composition) is contacted with a cell or tissue, or administered to a subject in need thereof, in an amount and for a time sufficient to modulate (increase or decrease) one or more of the following parameters: adaptor functions (e.g., homologous or non-homologous adaptor functions), e.g., rate, efficiency, robustness, and/or specificity of polypeptide chain initiation or extension; ribosome binding and/or occupancy; regulatory function (e.g., gene silencing or signaling); cell fate; mRNA stability; protein stability; protein transduction; protein compartmentalization. A parameter can be adjusted, e.g., by at least 5% (e.g., at least 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, or more) as compared to a reference tissue, cell, or subject (e.g., a healthy, wild-type, or control cell, tissue, or subject).

All references and publications cited herein are incorporated by reference.

The following examples are provided to further illustrate some embodiments of the present invention, but are not intended to limit the scope of the invention; it will be appreciated by their exemplary nature that other procedures, methods or techniques known to those skilled in the art may alternatively be used.

Examples of the invention

List of example contents:

example 1: production of TREM in mammalian production host cells by transient transfection

This example describes TREM produced in a mammalian host cell that transiently expresses TREM.

Plasmid production

To generate a plasmid comprising a sequence encoding TREM, in this example an iMet-CAT TREM, which is a DNA fragment containing one copy of sequence AGCAGAGTGGCGCAGCGGAAGCGTGCTGGGCCCATAACCCAGAGGTCGATGGATCGAAACCATCCTCTGCTA (SEQ ID NO:262), was synthesized and cloned into a pLKO.1-puro-mCherry backbone plasmid with the U6 promoter following the manufacturer's instructions and standard molecular cloning techniques.

Transfection

The above three (3) μ g plasmids were used to transfect T175 flasks of HEK293T cells plated at 80% confluence using 9uL lipofectamine RNAiMax reagent according to the manufacturer's instructions. Cells were harvested 48 hours post transfection for purification.

Purification Using Small RNA isolation kit

Cells overexpressing met are lysed. To generate small RNA (srna) fractions, RNA of less than 200 nucleotides is isolated from the remainder of the total RNA pool in the lysate using a small RNA isolation kit (e.g. Qiagen miRNeasy kit) according to the manufacturer's instructions. To further exclude larger RNAs, LiCl precipitation was performed to remove the remaining large RNAs in the sRNA fraction. Finally, the sRNA fraction was added to a G50 column to remove RNA of less than 10 nucleotides from the sRNA fraction and to perform buffer exchange.

To isolate TREM from sRNA fractions, a probe binding method was used. The iMet-CAT-TREM was bound and purified using a biotinylated capture probe corresponding to the DNA probe or 2'-OMe nucleic acid, which is complementary to a unique region of the target TREM being purified, in this example the probe binds biotin at the 5' end, sequence TAGCAGAGGATGGTTTCGATCCATCA (SEQ ID NO: 267). The sRNA fraction was incubated with annealing buffer and biotinylated capture probes at 90 ℃ for 4-5 minutes and cooled to 25 ℃ at a rate of 0.1 ℃/s.

The mixture was then incubated with binding buffer and streptavidin-conjugated rnase-free magnetic beads for 15 minutes to allow the DNA-TREM complex to bind to the beads. The mixture was then added to a magnetic separator stand and washed 2-3 times with wash buffer. The TREM retained on the beads is eluted by adding an elution buffer with or without dnase to ensure complete removal of the DNA capture probe, and then mixed with a pharmaceutically acceptable excipient to prepare the test TREM product.

Example 2: production of TREM in mammalian production host cells from stable cell lines

This example describes the manufacture of TREM produced in a mammalian host cell that stably expresses TREM.

Preparation of TREM-expressing lentivirus

To prepare TREM-expressing lentiviruses in 10mm dishes, packaging cells, such as HEK293T cells (293T cells)Forward transfection was performed using TransIT-LT1 transfection reagent according to the manufacturer's instructions with the following: mu.g of plasmid comprising the sequence encoding TREM as described in example 1 and 9. mu.g of ViraPower lentivirus packaging mixture.

After 18 hours, the medium was changed to fresh high FBS (30% FBS) medium without antibiotics, and after 24 hours, the virus-containing medium was collected and stored at 4 ℃. An additional 15mL of high FBS medium was added to the plates and harvested after 24 hours. The two virus-containing media harvests were combined and filtered through a 0.45 micron filter. Viral copy number was assessed using the Lenti-XqRT-PCR titration kit according to the manufacturer's protocol.

Transduction of host cells with a lentivirus expressing TREM

To transduce cells with a lentivirus expressing TREM, the lentivirus-containing medium was diluted 1:4 with complete cell culture medium in the presence of 10. mu.g/mL polybrene and added to the cells. In this example, 293T cells were used. The plates were spun at 1000Xg for 2 hours to spin-infect the cells. After 18 hours, the medium was changed to allow the cells to recover. Forty-eight hours after transduction, puromycin (2. mu.g/mL) antibiotic selection was performed for 5-7 days with the untransduced control cell population.

TREM was isolated, purified and formulated as described in example 1 to produce a TREM preparation.

Purification using phenol chloroform extraction

Total RNA pools of cells were recovered from the cells by guanidinium thiocyanate-phenol-chloroform extraction and concentrated by ethanol precipitation as described in J.Sambrook and D.Russell (2001) Molecular Cloning: A Laboratory Manual [ Molecular Cloning: A Laboratory Manual ], Vol.2, Cold Spring Harbor Laboratory Press [ Cold Spring Harbor Laboratory Press ], N.Y., U.S. 3 rd edition 2. And then by a method described in e.g. Cathala, g. et al, DNA, 1983; 329-35 high lithium salt precipitation will precipitate total tRNA pools from larger nucleic acids (including rRNA and DNA). The eluted fraction containing TREM is further purified by probe binding.

The TREM fraction is incubated with an annealing buffer and a biotinylated capture probe corresponding to the DNA probe or 2' -OMe nucleic acid, which is complementary to a unique region of the target TREM to be purified. In this example, a TREM containing iMet-CAT was purified using a probe conjugated at the 5' end to biotin (having sequence TAGCAGAGGATGGTTTCGATCCATCA (SEQ ID NO: 267)). The mixture was incubated at 90 ℃ for 4-5 minutes and then cooled to 25 ℃ at a rate of 0.1 ℃/s.

The mixture was then incubated with binding buffer and streptavidin-conjugated rnase-free magnetic beads for 15 minutes to allow the DNA-TREM complex to bind to the beads. The mixture was then added to a magnetic separator stand and washed 2-3 times. The TREM retained on the beads is eluted by adding an elution buffer with or without dnase to ensure complete removal of the DNA capture probe, and then mixed with a pharmaceutically acceptable excipient to prepare the test TREM product.

Example 3: production of TREM in mammalian production host cells from stable cell lines

This example describes the production of TREM from crude cell lysates produced from mammalian host cells.

Generation of Stable cells expressing TREM

In this example, a plasmid comprising a sequence encoding TREM was generated as described in example 1 or 2. Preparation of TREM expressing lentivirus and transduction of host cells with TREM expressing lentivirus was performed as described in example 2.

Purification from crude cell lysate

Cells overexpressing TREM, in this example iMet-CAT-TREM, are lysed and the lysed material is incubated with an annealing buffer and a biotinylated capture probe corresponding to the DNA probe or 2' -OMe nucleic acid, which is complementary to a unique region of the target TREM to be purified. In this example, a TREM containing iMet-CAT was purified using a probe conjugated at the 5' end to biotin (having sequence TAGCAGAGGATGGTTTCGATCCATCA (SEQ ID NO: 267)). The mixture was incubated at 90 ℃ for 4-5 minutes and then cooled to 25 ℃ at a rate of 0.1 ℃/s.

The mixture was then incubated with binding buffer and streptavidin-conjugated rnase-free magnetic beads for 15 minutes to allow the DNA-TREM complex to bind to the beads. The mixture was then added to a magnetic separator stand and washed 2-3 times. The TREM retained on the beads is eluted by adding an elution buffer with or without dnase to ensure complete removal of the DNA capture probe, and then mixed with a pharmaceutically acceptable excipient to prepare the test TREM product.

Example 4: delivery of TREM to mammalian cells

This example describes the delivery of TREM to mammalian cells.

To ensure correct folding, TREM was heated at 85 ℃ for 2 minutes and then rapidly cooled at 4 ℃ for 5 minutes. For delivery of TREM to mammalian cells, the RNAiMax reagent was used according to the manufacturer's instructions, at U2OS (U-2 OS: (R) (R))HTB-96^TM)、H1299(NCI-H1299(CRL-5803^TM) And HeLa (HeLa: (C))CCL-2^TM) 100nM of two TREM preparations labeled with Cy3 at different locations (Cy3-iMET-1 and Cy3-iMET-2)) were transfected into the cells. After 18 hours, the transfection medium was removed and replaced with fresh complete medium (U2 OS: McCoy's 5A, 10% FBS, 1% PenStrep; H1299: RPMI1640, 10% FBS, 1% PenStrep; HeLa: EMEM, 10% FBS, 1% PenStrep).

To observe TREM delivery to cells, cells are monitored in a living cell assay system. In this example, IncuCyte (from Essen BioScience) was used to monitor cells. Cells were monitored for 4 days (20x, red 550 ms).

The Cy3 fluorescent signal was readily detected from cells that had delivered Cy 3-labeled TREM. Cy3 fluorescence signals were observed for more than 48 hours from cells that had delivered TREM. Detection of Cy-3 fluorescence from the cells confirmed the delivery of Cy 3-labeled TREM to the cells.

Example 5: increased cell growth in mammalian cells using TREM

This example describes the increase in cell growth of mammalian cells following TREM delivery.

To ensure correct folding, the iMet TREM was heated at 85 ℃ for 2 minutes and then rapidly cooled at 4 ℃ for 5 minutes. For delivery of iMet TREM to mammalsBiological cells, using the RNAiMax reagent according to the manufacturer's instructions, in U2OS (U-2 OS: (B) (B))HTB-96^TM)、H1299(NCI-H1299(CRL-5803^TM) And HeLa (HeLa: (C))CCL-2^TM) 100nM Cy 3-labeled iMet TREM was transfected into the cells. As a control, non-targeted control siRNA labeled with Cy3 was delivered to cells. After 18 hours, the transfection medium was removed and replaced with fresh complete medium (U2 OS: McCoy's 5A, 10% FBS, 1% PenStrep; H1299: RPMI1640, 10% FBS, 1% PenStrep; HeLa: EMEM, 10% FBS, 1% PenStrep). To observe changes in cell growth, cells were monitored in a live cell assay system (in this example, IncuCyte (from eisen biosciences)) for 4 days (20x, phase contrast).

Delivery of the met TREM to U2OS cells (fig. 1A), H1299 (fig. 1B) or Hela cells (fig. 1C) resulted in a significant increase in cell growth in all cell lines tested. The increase in cell growth was compared to that observed by delivery of a Cy 3-labeled non-targeted control (Cy 3-NTC). The data indicate that delivery of TREM to cells results in increased proliferation and growth.

Example 6: determination of TREM translational Activity in human cell extract cell-free protein Synthesis (hCFPS) lysates

This example describes TREM-mediated increase in translational activity in a cell-free lysate system.

Preparation of human cell extract

HEK293T cells were grown to about 80% confluence in 40X 150mm dishes. Cells were harvested, washed in PBS, resuspended in ice-cold hypotonic lysis buffer (20mM HEPES pH 7.6, 10mM KAc, 1.5mM MgAc, 5mM DTT, and 5X complete EDTA-free protease inhibitor cocktail) at a ratio of 1:1 and incubated on ice for 30 minutes. Cells were lysed using a Dounce homogenizer or by passing the lysate through a 27G needle until > 95% of the cells were destroyed. The lysate was centrifuged at 14,000g for 10 minutes at 4 ℃, and the supernatant was collected and diluted with hypotonic lysis buffer to obtain a protein solution of about 15 mg/ml.

Transcribing mRNA

The mRNA transcription template was designed with a T7 polymerase promoter, a beta-globin 3 'UTR, a nanoLuc ORF, and a short artificial 3' UTR. The template was amplified by PCR and the capped and polyadenylated mRNA was transcribed using the tailed histribe T7 ARCA mRNA kit (new england biosciences) according to the manufacturer's recommended protocol.

TREM translational activity assay in hCFPS lysates

Translation reactions were established in translation buffer (16mM HEPES pH 7.6, 2.2mM MgAc, 60mM KCl, 0.02mM whole amino acid mixture, 1mM ATP, 0.5mM GTP, 20mM creatine phosphate, 0.1. mu.g/. mu.L creatine kinase, 0.1mM spermidine, 2U/. mu.l RiboLock RNase inhibitor) containing 35% HEK293T lysate, 0.02. mu.M capped and polyadenylated nanoLuc mRNA and 2. mu.M cell purified TREM (purified according to example 2). The reaction was performed in triplicate at 10. mu.l for 30 min at 37 ℃. For the control reactions, one control reaction was performed without adding TREM to the reaction, and one control reaction was performed without adding mRNA to the reaction. The NanoLuc activity was then detected by mixing each reaction with 40 μ l of room temperature Nano-Glo luciferase assay system (Promega) and reading the luminescence in a plate reader.

As shown in fig. 2, the iMET TREM reaction resulted in about a 1.5-fold increase in NanoLuc expression compared to the control reaction (buffer). The data show that delivery of TREM results in increased translation of nanoLuc mRNA as reflected by increased luminescence.

Example 7: production of TREM in mammalian production host cells and its use in modulating cellular function-1

This example describes the manufacture of TREM produced in a mammalian host cell.

Plasmid production

To generate a plasmid comprising a sequence encoding TREM, in this example, iMet-CAT TREM, a DNA fragment having genomic position 6p22.2 and sequence AGCAGAGTGGCGCAGCGGAAGCGTGCTGGGCCCATAACCCAGAGGTCGATGGATCGAAACCATCCTCTGCTA (SEQ ID NO:622) was PCR amplified from human genomic DNA using the following primer pairs: 5 '-TGAGTTGGCAACCTGTGGTA (SEQ ID NO:623) and 5' -TTGGGTGTCCATGAAAATCA (SEQ ID NO: 624). This fragment was cloned into a plko.1puro backbone plasmid with the U6 promoter (or any other RNA polymerase III recruiting promoter) according to the manufacturer's instructions.

Transfection

One (1) mg of the above plasmid was used at 1X 10⁵Individual cells/mL were transfected into 1L cultures of HEK293T cells (Freestyle 293-F cells) in suspension. Cells were harvested at 24, 48, 72 or 96 hours post transfection to determine the optimal time point for TREM expression as determined by northern blot or quantitative PCR (q-PCR).

Purification of

At the optimized harvested cell density point, TREM was purified as previously described in Cayama et al, Nucleic Acids Research [ Nucleic Acids ]28(12), e64 (2000). Briefly, short RNAs (e.g., trnas) are recovered from cells by phenol extraction and concentrated by ethanol precipitation. The total tRNA in the precipitate was then separated from the larger nucleic acids (including rRNA and DNA) by stepwise isopropanol precipitation under high salt conditions. The eluted fraction containing TREM is further purified by probe binding. The TREM fraction is incubated with annealing buffer and iMet-CAT-TREM is purified using a biotinylated capture probe corresponding to the DNA probe or 2'-OMe nucleic acid, which is complementary to a unique region of the target TREM being purified, in this example the probe binds biotin at the 3' end, sequence UAGCAGAGGAUGGUUUCGAUCCAUCA (SEQ ID NO: 625). The mixture was incubated at 90 ℃ for 2-3 minutes, then rapidly cooled to 45 ℃ and incubated overnight at 45 ℃. The mixture was then incubated with binding buffer and streptavidin-conjugated magnetic beads without rnase for 3 hours, previously heated to 45 ℃, to bind the DNA-TREM complex to the beads. The mixture was then added to the pre-equilibrated column in the magnetic separator stand and washed 4 times. The TREM product is tested by adding elution buffer pre-warmed to 80 ℃ and then mixed with pharmaceutically acceptable excipients, and the TREM retained on the beads is eluted three times.

Use of

One microgram of the test TREM preparation and control agent is contacted with a cultured cell line (e.g., HEP-3B or HEK293T), tissue, or subject by transfection, electroporation, or liposome delivery for a time sufficient for the TREM preparation to modulate translation levels or activity of the cells relative to the control agent.

Example 8: production of TREM in mammalian production host cells and its use in modulating cellular function-2

This example describes the manufacture of TREM produced in a mammalian host cell.

Plasmid production

To generate a plasmid comprising a sequence encoding TREM, in this example an iMet-CAT-TREM (which is a DNA fragment comprising at least one copy of sequence AGCAGAGTGGCGCAGCGGAAGCGTGCTGGGCCCATAACCCAGAGGTCGATGGATCGAAACCATCCTCTGCTA (SEQ ID NO: 626)) was synthesized and cloned into a pLKO.1puro backbone plasmid with the U6 promoter (or any other RNA polymerase III recruited promoter) following the manufacturer's instructions and standard molecular cloning techniques.

Transfection

One (1) mg of the above plasmid was used at 1X 10⁵Individual cells/mL were transfected into 1L cultures of HEK293T cells (Freestyle 293-F cells) in suspension. Cells were harvested 24, 48, 72 or 96 hours post transfection to determine the optimal time point for TREM expression as determined by northern blot or quantitative PCR (q-PCR) or nanopore sequencing.

Purification of

At the optimized harvest time point, cells were lysed and separated from lysates of less than 200 nucleotides of RNA using a small RNA isolation kit according to the manufacturer's instructions to produce small RNA (srna) fractions.

To prepare the affinity purification reagents, streptavidin-conjugated rnase-free magnetic beads were incubated with 200mM biotinylated oligonucleotide (which corresponds to a DNA probe or 2' -OMe nucleic acid complementary to the target TREM being purified) for 30 minutes at room temperature. In this example, a probe having the sequence 5' biotin-TAGCAGAGGATGGTTTCGATCCATCA (SEQ ID NO:627) was used to purify-iMet-CAT-TREM. The beads were washed and heated at 75 ℃ for 10 minutes.

The sRNA fraction was heated at 75 ℃ for 10 minutes and then mixed with the above-described affinity purification reagents. The mixture was incubated at room temperature for 3 hours to allow TREM to bind in a sequence specific manner to the bead-bound DNA probes. The beads were then washed until the absorbance of the wash solution at 260nm was close to zero. Alternatively, the beads are washed three times and the final wash is checked by UV spectroscopy to measure the amount of nucleic acid present in the final wash. TREM retained on beads was eluted three times using rnase-free water, which can be preheated to 80 ℃, and then mixed with a pharmaceutically acceptable excipient to prepare test TREM products.

Use of

One microgram of the test TREM preparation and control agent is contacted with a cultured cell line (e.g., HeLa, HEP-3B, or HEK293T), tissue, or subject by transfection, electroporation, or liposome delivery for a time sufficient for the TREM preparation to modulate translation levels or activity of the cell relative to the control agent.

Example 9: production of TREM in modified mammalian production host cells expressing oncogenes

This example describes the production of TREM in mammalian host cells to overexpress Myc.

Plasmid production and host cell modification

To make the production host cells of this example, HeLa cells (A) were prepared using conventional molecular biology techniquesCCL-2^TM) Or HEP-3B cells (HB-8064^TM) Transfection with a plasmid containing the gene sequence encoding the c-myc oncogene protein (e.g., pcDNA3-cmyc (Addgene plasmid # 16011)). The resulting cell line is referred to herein as a HeLamyc + host cell or a HEP-3Bmyc + host cell.

Preparation of TREM-expressing lentivirus

To prepare a lentivirus expressing TREM, HEK293T cells were co-transfected with Lipofectamine2000 according to the manufacturer's instructions using: mu.g of each packaging vector (pRSV-Rev, pCMV-VSVG-G and pCgpV) and 9. mu.g of plasmid comprising a sequence encoding TREM as described in example 7. After 24 hours, the medium was changed to fresh antibiotic-free medium, and after 48 hours, the virus-containing supernatant was collected and centrifuged at 2000rpm for 10 minutes, and then filtered through a 0.45 μm filter.

Transduction of host cells with a lentivirus expressing TREM

100,000 HeLamyc + or HEP-3Bmyc + host cells were transduced with two (2) mL of virus prepared as described above in the presence of 8. mu.g/mL of polybrene. Forty-eight hours after transduction, puromycin (2. mu.g/mL) antibiotic selection was performed for 2-7 days with the untransduced control cell population.

TREM is isolated, purified and formulated as described in example 7 or 8 to produce a TREM composition or preparation.

Example 10: production of TREM-producing host cells modified to inhibit a repressor of tRNA synthesis

This example describes the preparation of Hek293Maf-/TRM1 cells for the production of TREM.

Maf1 is a repressor of tRNA synthesis. The Maf1 knockout HEK293T cell line was generated using standard CRISPR/Cas knockout techniques, for example, CRISPR/Cas systems can be designed to introduce frame shift mutations in the coding exon of Maf1 to reduce expression of Maf1 or to knock out expression of Maf1 to generate HEK293 Maf-cell lines with reduced expression levels and/or activity of Maf 1. This cell line is then transfected with an expression plasmid for the modification enzyme Trm1(tRNA (guanine 26-N2) -dimethyltransferase), e.g., pCMV6-XL4-Trm1, and selected with a selection marker, e.g., neomycin, to generate a stable cell line overexpressing Trm1 (Hek293Maf-/TRM1 cells).

Hek293Maf-/TRM1 cells can be used as production host cells to produce TREM as described in any of examples 7-9.

Example 11: production of TREM in modified mammalian production host cells overexpressing oncogenes and tRNA modifying enzymes

This example describes the production of TREM in mammalian host cells to overexpress Myc and Trm 1.

Plasmid production

In this example, a TREM-containing plasmid was generated as described in example 7 or 8.

Host cell modification, transduction and purification

By converting a retrovirus expressing a myc oncogene from pBABEpuro-c-myc^T58ATransduction of the plasmid into HEK293T cells can result in human cell lines stably overexpressing Myc oncogenes, such as HEK 293T. To generate a retrovirus expressing myc, the calcium phosphate approach was used with the human c-myc retroviral vector pBABEpuro-c-myc^T58AAnd packaging vector psi 2 vector to transfect HEK293T cell. After 6 hours, the transfection medium was removed and replaced with fresh medium. After 24 hours of incubation, the medium was collected and filtered through a 0.45um filter. For retroviral infection, HEK293T cells were infected with retrovirus and polybrene (8ug/ml) and spun at 2500rpm for 1 hour at 18 ℃. After 24 hours, the cell culture medium was replaced with fresh medium and after 24 hours, the cells were selected with 2 μ g/mL puromycin. Once cells stably overexpressing the oncogene Myc were established, they were transfected with a Trm1 plasmid (e.g., pCMV6-XL4-Trm1 plasmid) and selected using a selection marker (neomycin in this case) to generate a stable cell line overexpressing Trm1 in addition to Myc. Meanwhile, HEK293T cells and PLKO.1-TREM vectors were used to generate lentiviruses overexpressing TREM as described in example 9.

One hundred thousand (1X 10) were transduced with TREM virus in the presence of 8. mu.g/mL polybrene⁵) And (3) a cell overexpressing Myc and Trm 1. The medium was changed after 24 hours. Forty-eight hours after transduction, antibiotic selection was performed using 2 μ g/mL puromycin with an untransduced control cell population for 2-7 days. TREM was isolated, purified and formulated using the methods described in examples 7 or 8 to produce TREM preparations.

Example 12: TREM generation of error load

This example describes the generation of a TREM loaded with an amino acid different from its native anti-codon.

TREM was produced as described in any of examples 7-11. The TREM product is loaded with heterologous amino acids using in vitro loading reactions known in the art (see, e.g., Walker & Fredrick (2008) Methods [ Methods ] (san diego, ca) 44(2): 81-6). Briefly, a purified TREM, e.g., a TREM comprising tRNA-val (gtg), is placed in a buffer with a heterologous amino acid of interest (e.g., glutamic acid) and a corresponding aminoacyl-tRNA synthetase (e.g., Valyl-tRNA synthetase that is mutated to enhance tRNA misloading) to induce TREM loading.

For isolation of aminoacyl-TREM, the in vitro loading reaction is passed through a spin column and determined on the basis of A ₂₆₀The concentration of absorbance and the degree of aminoacylation using acid gel electrophoresis. Aminoacylated TREM can also be isolated by: binding to His 6-tagged EF-Tu ("His 6", published as SEQ ID NO:628), followed by affinity chromatography on Ni-NTA agarose, phenol-chloroform extraction and subsequent nucleic acid precipitation, e.g., Rezgui et al, 2013, PNAS [ Proc. Natl. Acad. Sci. USA)]110:12289 and 12294.

Example 13: generation of TREM fragments (in vitro)

This example describes the in vitro production of TREM fragments from TREM produced in mammalian host cells.

TREM is prepared as described in any of examples 7-13 above. Enzymatic cleavage assays using enzymes known to produce tRNA fragments (e.g., rnase a or angiogenin) are used to produce the fragments for administration to cells, tissues, or subjects.

Briefly, TREM made as described above is incubated in one of the following: 0.1MHepes/NaOH, pH 7.4, 10nM RNase A final concentration, 10 minutes at 30 ℃ or 0.1M MES, 0.1M NaCl, pH 6.0, effective amounts of angiogenin, and BSA at 37 ℃ for 6 hours.

To isolate the TREM fragment of interest after enzymatic treatment, a sequence affinity purification procedure is performed, as described above.

Example 14: production of TREM fragments in a cell expression System

This example describes the generation of TREM fragments in a cell expression system.

A cell line stably overexpressing TREM is generated as described in any one of examples 7-9 or 11. Hek293T cells overexpressing TREM were treated with 0.5. mu.g/ml recombinant angiogenin for 90 minutes, and then total RNA was extracted with Trizol. RNA of less than 200 nucleotides was size-selected using a small RNA isolation kit according to the manufacturer's instructions. Streptavidin-conjugated rnase-free magnetic beads were incubated with 200mM biotinylated oligonucleotides (which correspond to probes complementary to unique regions of the TREM fragment to be purified or DNA probes) for 30 minutes at room temperature. The beads were washed and heated at 75 ℃ for 10 minutes. The size-selected RNA eluate was also heated at 75 ℃ for 10 minutes and then mixed with the beads. The TREM-bead mixture was incubated at room temperature for 3 hours to allow binding of TREM to the bead-bound DNA probes. The beads were then washed until the wash solution at 260nm was near zero (0). Alternatively, the beads are washed three times and the final wash is checked by UV spectroscopy to measure the amount of nucleic acid present in the final wash. TREM retained on the beads was eluted 3 times using RNase-free water pre-heated to 80 ℃ or elution buffer pre-heated to 80 ℃.

Example 15: TREM translational activity assay

This example describes an assay to assess the ability of TREM to incorporate nascent polypeptide chains.

Translation of FLAG-AA-His peptide sequences

The test TREM was assayed in an in vitro translation reaction using mRNA encoding the peptide FLAG-XXX-His6x ("His 6", disclosed as SEQ ID NO:628), where XXX is 3 consecutive codons corresponding to the anti-codon of the test TREM.

tRNA depleted rabbit reticulocyte lysate (Jackson et al 2001.RNA7:765-773) was incubated for 1 hour at 30 ℃ with 10-25ug/mL of test TREM and 10-25ug/mL of tRNA required for FLAG and His tag translation. In this example, the TREM used was Ile-GAT-TREM, so the peptide used was FLAG-LLL-His6x ("His 6", disclosed as SEQ ID NO:628) and the TREM added was TREM-Ile-GAT, and the following (which was added to translate the peptide FLAG and HIS tags): tRNA-Asp-GAC, tRNA-Tyr-TAC, tRNA-Lys-AAA, tRNA-Lys-AAAG, tRNA-Asp-GAT and tRNA-His-CAT. To determine whether the test TREM was functionally incorporated into the nascent peptide, an ELISA capture assay was performed. Briefly, an immobilized anti-His 6X antibody ("His 6", disclosed as SEQ ID NO:628) was used to capture the FLAG-LLL-His6x peptide ("His 6", disclosed as SEQ ID NO:628) from the reaction mixture. The reaction mixture is then washed away and the peptides are detected with enzyme-conjugated anti-FLAG antibody, which reacts with the substrate in the ELISA detection step. If the TREM produced is functional, the FLAG-LLL-His6 peptide ("His 6", disclosed as SEQ ID NO:628) is produced and detected by an ELISA capture assay.

If the TREM produced is non-functional, the FLAG-LLL-His6 peptide ("His 6", disclosed as SEQ ID NO:628) is not produced and is not detectable by ELISA capture assay.

Translation inhibition assay

This assay describes a test TREM that has the function of a translation adaptor by rescuing inhibitory mutations and allowing translation of the complete protein. A test TREM, in this example Ile-CUA-TREM, is generated such that it comprises the sequence of the Ile-GAT-TREM body, but has an anticodon sequence corresponding to CUA rather than GAT. HeLa cells were co-transfected as described in Geslain et al 2010.J Mol Biol. [ J. Mol. Biol. 396: 821-831): 50ng TREM and 200ng DNA plasmid encoding mutant GFP (containing a TAG stop codon at position S29). HeLa cells transfected with GFP plasmid alone served as negative controls. After 24 hours, cells were harvested and analyzed for fluorescence recovery by flow cytometry. Fluorescence was read as an emission peak at 509nm (excitation at 395 nm). It is expected that if the test TREM is functional, it may or will be sufficient to rescue the terminating mutation in the GFP molecule and produce a full-length fluorescent protein, which is detected by flow cytometry. If the test TREM is not or poorly functional, the terminating mutation may not be rescued and the GFP molecule does not fluoresce, so flow cytometry detects reduced or no GFP signal.

In vitro translation assay

This assay describes a test TREM with translational adaptor molecule function by successful incorporation into nascent polypeptide chains in an in vitro translation reaction. First, rabbit reticulocytes depleted of endogenous tRNA using antisense or complementary oligonucleotides are generatedA lysate, said antisense or complementary oligonucleotide (i) targeting a sequence between an anticodon and a variable loop; or (ii) binds to the region between the anti-codon and the variable loop (see, e.g., Cui et al 2018.Nucleic Acids Res [ Nucleic acid research ]].46(12):6387-6400). In addition to 2ug/uL of GFP-encoding mRNA, 10-25ug/mL of test TREM was added to the depleted lysate. Non-depleted lysates containing GFP mRNA (whether or not test TREM was added) were used as positive controls. Depleted lysates containing GFP mRNA but no test TREM added were used as negative controls. Using λ_ex485/λ_emThe progress of the translation of GFP mRNA was monitored at 37 ℃ for 3-5 hours by an increase in fluorescence on the microplate reader 528. The experimental sample is expected to produce a similar level of fluorescence over time as the positive control, and a higher level of fluorescence over time as compared to the negative control. If so, these results may indicate that the test TREM is sufficient or can replenish depleted lysate and therefore may be functional.

Example 16: determining modulation of cellular status

This example describes assays for detecting TREM activity in modulating cell status (e.g., cell death).

TREM fragments were generated as described in example 13. One (1) uM TREM fragment was transfected into HEK293T cells using Lipofectamine 3000 and incubated at one hour intervals for 1-6 hours before cell lysis. Cell lysates were analyzed by western blot and blots probed with antibodies against sum cleaved caspases 3 and 9 as a readout for apoptosis. To measure cell viability, cells were washed and fixed with 4% paraformaldehyde in PBS for 15 minutes at room temperature. The fixed and washed cells were then treated with 0.1% Triton X-100 for 10 minutes at room temperature and washed three times with PBS. Finally, the cells were treated with TUNEL assay reaction mixture for 1 hour at 37 ℃ in the dark. Samples were analyzed by flow cytometry.

Example 17: determination of unsupported TREM Activity to modulate autophagy

This example describes an assay to test the ability of unloaded TREM to modulate, e.g., induce autophagy, e.g., activate GCN 2-dependent stress response (starvation) pathway signaling, inhibit mTOR, or activate autophagy.

The test unloaded trem (utrem) preparation was delivered to HEK293T or HeLa cells by transfection or liposome delivery. After delivery of the uTREM, a time course ranging from 30 minutes to 6 hours will be performed, including the interval time points of one hour. The cells were then trypsinized, washed and lysed. The same procedure was performed using a loaded control TREM and random RNA oligonucleotides as controls. Cell lysates were analyzed by western blotting and blotted with antibodies directed against known readings of GCN2 pathway activation, mTOR pathway inhibition, or autophagy induction, including but not limited to phospho-eIF 2a, ATF4, phospho-ULK 1, phospho-4 EBP1, phospho-eIF 2a, phospho-Akt, and phospho-p 70S 6K. Total protein loading controls (e.g., GAPDH, actin, or tubulin, and unmodified (i.e., non-phosphorylated) signaling proteins (i.e., controls using eIF2a as phospho-eIF 2 a)) were probed as loading controls. Delivery of uTREM is or can be expected to show activation of the GCN2 starvation signaling pathway, autophagy pathway, and/or inhibition of the mTOR pathway, as compared to a control, as determined by western blot analysis.

Example 18: determination of the Activity of unsupported TREM (mTREM)

This example describes an assay to test the function of mTREM produced in a cellular system using plasmid transfection followed by in vitro error loading.

In this example, mTREM may translate mutant mRNA into wild-type (WT) protein by incorporating WT amino acids into the protein, although the mRNA contains mutant codons. This example uses GFP mRNA molecules with mutations of T203I or E222G, which block GFP excitation at wavelengths of 470nm and 390nm, respectively. GFP mutants that block GFP fluorescence can also be used as reporter proteins in this assay. Briefly, using an in vitro translation assay, rabbit reticulocyte lysate containing the GFP E222G mutant mRNA (GAG → GGG mutation) and excess mTREM (in this case Glu-CCC-TREM) was used. As a negative control, TREM with no error load was added to the reaction. If mTREM is functional, GFP protein will or can be expected to fluoresce when illuminated with a fluorometer at 390nm excitation wavelength. If mTREM is not or is less functional, the GFP protein will only fluoresce when excited at 470nm, as observed in the negative control.

Sequence listing

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<213> Intelligent people

<400> 4

ggggaattag ctcaagtggt agagcgcttg cttagcatgc aagaggtagt gggatcgatg 60

cccacattct cca 73

<210> 5

<211> 73

<212> DNA

<213> Intelligent people

<400> 5

ggggaattag ctcaagcggt agagcgcttg cttagcatgc aagaggtagt gggatcgatg 60

cccacattct cca 73

<210> 6

<211> 73

<212> DNA

<213> Intelligent people

<400> 6

ggggaattag ctcaagtggt agagcgcttg cttagcatgc aagaggtagt gggatcaatg 60

cccacattct cca 73

<210> 7

<211> 73

<212> DNA

<213> Intelligent people

<400> 7

ggggaattag ctcaagtggt agagcgctcg cttagcatgc gagaggtagt gggatcgatg 60

cccgcattct cca 73

<210> 8

<211> 73

<212> DNA

<213> Intelligent people

<400> 8

ggggaattag cccaagtggt agagcgcttg cttagcatgc aagaggtagt gggatcgatg 60

cccacattct cca 73

<210> 9

<211> 72

<212> DNA

<213> Intelligent people

<400> 9

gggggtgtag ctcagtggta gagcgcgtgc ttagcatgca cgaggccccg ggttcaatcc 60

ccggcacctc ca 72

<210> 10

<211> 72

<212> DNA

<213> Intelligent people

<400> 10

gggggtgtag ctcagtggta gagcgcgtgc ttagcatgta cgaggtcccg ggttcaatcc 60

ccggcacctc ca 72

<210> 11

<211> 72

<212> DNA

<213> Intelligent people

<400> 11

ggggatgtag ctcagtggta gagcgcatgc ttagcatgca tgaggtcccg ggttcgatcc 60

ccagcatctc ca 72

<210> 12

<211> 72

<212> DNA

<213> Intelligent people

<400> 12

gggggtgtag ctcagtggta gagcgcgtgc ttagcatgca cgaggccctg ggttcaatcc 60

ccagcacctc ca 72

<210> 13

<211> 72

<212> DNA

<213> Intelligent people

<400> 13

gggggtatag ctcagcggta gagcgcgtgc ttagcatgca cgaggtcctg ggttcaatcc 60

ccaatacctc ca 72

<210> 14

<211> 72

<212> DNA

<213> Intelligent people

<400> 14

gggggtgtag ctcagtggta gagcgcgtgc ttagcatgca cgaggccccg ggttcaatcc 60

ctggcacctc ca 72

<210> 15

<211> 73

<212> DNA

<213> Intelligent people

<400> 15

gggggattag ctcaaatggt agagcgctcg cttagcatgc gagaggtagc gggatcgatg 60

cccgcatcct cca 73

<210> 16

<211> 73

<212> DNA

<213> Intelligent people

<400> 16

ggggaattag ctcaggcggt agagcgctcg cttagcatgc gagaggtagc gggatcgacg 60

cccgcattct cca 73

<210> 17

<211> 72

<212> DNA

<213> Intelligent people

<400> 17

ggggatgtag ctcagtggta gagcgcatgc ttcgcatgta tgaggtcccg ggttcgatcc 60

ccggcatctc ca 72

<210> 18

<211> 72

<212> DNA

<213> Intelligent people

<400> 18

ggggatgtag ctcagtggta gagcgcatgc ttcgcatgta tgaggccccg ggttcgatcc 60

ccggcatctc ca 72

<210> 19

<211> 72

<212> DNA

<213> Intelligent people

<400> 19

ggggatgtag ctcagtggta gagcgcgcgc ttcgcatgtg tgaggtcccg ggttcaatcc 60

ccggcatctc ca 72

<210> 20

<211> 72

<212> DNA

<213> Intelligent people

<400> 20

gggggtgtag ctcagtggta gagcgcgtgc ttcgcatgta cgaggccccg ggttcgaccc 60

ccggctcctc ca 72

<210> 21

<211> 72

<212> DNA

<213> Intelligent people

<400> 21

gggggtgtag ctcagtggta gagcgcatgc tttgcatgta tgaggtcccg ggttcgatcc 60

ccggcacctc ca 72

<210> 22

<211> 72

<212> DNA

<213> Intelligent people

<400> 22

ggggatgtag ctcagtggta gagcgcatgc tttgcatgta tgaggtcccg ggttcgatcc 60

ccggcatctc ca 72

<210> 23

<211> 72

<212> DNA

<213> Intelligent people

<400> 23

ggggatgtag ctcagtggta gagcgcatgc tttgcatgta tgaggccccg ggttcgatcc 60

ccggcatctc ca 72

<210> 24

<211> 72

<212> DNA

<213> Intelligent people

<400> 24

ggggatgtag ctcagtggta gagcgcatgc tttgcacgta tgaggccccg ggttcaatcc 60

ccggcatctc ca 72

<210> 25

<211> 72

<212> DNA

<213> Intelligent people

<400> 25

gggggtgtag ctcagtggta gagcgcatgc tttgcatgta tgaggcctcg ggttcgatcc 60

ccgacacctc ca 72

<210> 26

<211> 72

<212> DNA

<213> Intelligent people

<400> 26

gggggtgtag ctcagtggta gagcacatgc tttgcatgtg tgaggccccg ggttcgatcc 60

ccggcacctc ca 72

<210> 27

<211> 71

<212> DNA

<213> Intelligent people

<400> 27

gggggtgtag ctcagtggta gagcgcatgc tttgcatgta tgaggcctcg gttcgatccc 60

cgacacctcc a 71

<210> 28

<211> 73

<212> DNA

<213> Intelligent people

<400> 28

gggccagtgg cgcaatggat aacgcgtctg actacggatc agaagattcc aggttcgact 60

cctggctggc tcg 73

<210> 29

<211> 73

<212> DNA

<213> Intelligent people

<400> 29

gggccagtgg cgcaatggat aacgcgtctg actacggatc agaagattct aggttcgact 60

cctggctggc tcg 73

<210> 30

<211> 73

<212> DNA

<213> Intelligent people

<400> 30

ggccgcgtgg cctaatggat aaggcgtctg attccggatc agaagattga gggttcgagt 60

cccttcgtgg tcg 73

<210> 31

<211> 73

<212> DNA

<213> Intelligent people

<400> 31

gacccagtgg cctaatggat aaggcatcag cctccggagc tggggattgt gggttcgagt 60

cccatctggg tcg 73

<210> 32

<211> 73

<212> DNA

<213> Intelligent people

<400> 32

gccccagtgg cctaatggat aaggcactgg cctcctaagc cagggattgt gggttcgagt 60

cccacctggg gta 73

<210> 33

<211> 73

<212> DNA

<213> Intelligent people

<400> 33

gccccagtgg cctaatggat aaggcactgg cctcctaagc cagggattgt gggttcgagt 60

cccacctggg gtg 73

<210> 34

<211> 73

<212> DNA

<213> Intelligent people

<400> 34

gccccggtgg cctaatggat aaggcattgg cctcctaagc cagggattgt gggttcgagt 60

cccacccggg gta 73

<210> 35

<211> 73

<212> DNA

<213> Intelligent people

<400> 35

gccccagtgg cctaatggat aaggcattgg cctcctaagc cagggattgt gggttcgagt 60

cccatctggg gtg 73

<210> 36

<211> 73

<212> DNA

<213> Intelligent people

<400> 36

gccccagtgg cctgatggat aaggtactgg cctcctaagc cagggattgt gggttcgagt 60

tccacctggg gta 73

<210> 37

<211> 73

<212> DNA

<213> Intelligent people

<400> 37

ggccgcgtgg cctaatggat aaggcgtctg acttcggatc agaagattgc aggttcgagt 60

cctgccgcgg tcg 73

<210> 38

<211> 73

<212> DNA

<213> Intelligent people

<400> 38

gaccacgtgg cctaatggat aaggcgtctg acttcggatc agaagattga gggttcgaat 60

ccctccgtgg tta 73

<210> 39

<211> 73

<212> DNA

<213> Intelligent people

<400> 39

gaccgcgtgg cctaatggat aaggcgtctg acttcggatc agaagattga gggttcgagt 60

cccttcgtgg tcg 73

<210> 40

<211> 73

<212> DNA

<213> Intelligent people

<400> 40

gaccacgtgg cctaatggat aaggcgtctg acttcggatc agaagattga gggttcgaat 60

cccttcgtgg tta 73

<210> 41

<211> 73

<212> DNA

<213> Intelligent people

<400> 41

gaccacgtgg cctaatggat aaggcgtctg acttcggatc agaagattga gggttcgaat 60

cccttcgtgg ttg 73

<210> 42

<211> 73

<212> DNA

<213> Intelligent people

<400> 42

ggccgtgtgg cctaatggat aaggcgtctg acttcggatc aaaagattgc aggtttgagt 60

tctgccacgg tcg 73

<210> 43

<211> 85

<212> DNA

<213> Intelligent people

<400> 43

ggctccgtgg cgcaatggat agcgcattgg acttctagag gctgaaggca ttcaaaggtt 60

ccgggttcga gtcccggcgg agtcg 85

<210> 44

<211> 88

<212> DNA

<213> Intelligent people

<400> 44

ggctctgtgg cgcaatggat agcgcattgg acttctagtg acgaatagag caattcaaag 60

gttgtgggtt cgaatcccac cagagtcg 88

<210> 45

<211> 91

<212> DNA

<213> Intelligent people

<400> 45

ggctctgtgg cgcaatggat agcgcattgg acttctagct gagcctagtg tggtcattca 60

aaggttgtgg gttcgagtcc caccagagtc g 91

<210> 46

<211> 86

<212> DNA

<213> Intelligent people

<400> 46

ggctctgtgg cgcaatggat agcgcattgg acttctagat agttagagaa attcaaaggt 60

tgtgggttcg agtcccacca gagtcg 86

<210> 47

<211> 74

<212> DNA

<213> Intelligent people

<400> 47

gtctctgtgg cgcaatggac gagcgcgctg gacttctaat ccagaggttc cgggttcgag 60

tcccggcaga gatg 74

<210> 48

<211> 87

<212> DNA

<213> Intelligent people

<400> 48

ggctctgtgg cgcaatggat agcgcattgg acttctagcc taaatcaaga gattcaaagg 60

ttgcgggttc gagtccctcc agagtcg 87

<210> 49

<211> 74

<212> DNA

<213> Intelligent people

<400> 49

gtctctgtgg cgcaatcggt tagcgcgttc ggctgttaac cgaaaggttg gtggttcgat 60

cccacccagg gacg 74

<210> 50

<211> 74

<212> DNA

<213> Intelligent people

<400> 50

gtctctgtgg cgcaatcggc tagcgcgttt ggctgttaac taaaaggttg gcggttcgaa 60

cccacccaga ggcg 74

<210> 51

<211> 74

<212> DNA

<213> Intelligent people

<400> 51

gtctctgtgg tgcaatcggt tagcgcgttc cgctgttaac cgaaagcttg gtggttcgag 60

cccacccagg gatg 74

<210> 52

<211> 74

<212> DNA

<213> Intelligent people

<400> 52

gtctctgtgg cgcaatcggc tagcgcgttt ggctgttaac taaaaagttg gtggttcgaa 60

cacacccaga ggcg 74

<210> 53

<211> 74

<212> DNA

<213> Intelligent people

<400> 53

gtctctgtgg cgcaatcggt tagcgcgttc ggctgttaac cgaaaggttg gtggttcgag 60

cccacccagg gacg 74

<210> 54

<211> 74

<212> DNA

<213> Intelligent people

<400> 54

gtctctgtgg cgcaatcggt tagcgcattc ggctgttaac cgaaaggttg gtggttcgag 60

cccacccagg gacg 74

<210> 55

<211> 74

<212> DNA

<213> Intelligent people

<400> 55

gtctctgtgg cgcaatcggt tagcgcgttc ggctgttaac cgaaagattg gtggttcgag 60

cccacccagg gacg 74

<210> 56

<211> 74

<212> DNA

<213> Intelligent people

<400> 56

gtctctgtgg cgcaatcggt tagcgcgttc ggctgttaac tgaaaggttg gtggttcgag 60

cccacccagg gacg 74

<210> 57

<211> 74

<212> DNA

<213> Intelligent people

<400> 57

gtctctgtgg cgcaatgggt tagcgcgttc ggctgttaac cgaaaggttg gtggttcgag 60

cccatccagg gacg 74

<210> 58

<211> 74

<212> DNA

<213> Intelligent people

<400> 58

gtctctgtgg cgtagtcggt tagcgcgttc ggctgttaac cgaaaagttg gtggttcgag 60

cccacccagg aacg 74

<210> 59

<211> 74

<212> DNA

<213> Intelligent people

<400> 59

gtctctgtgg cgcaatcggc tagcgcgttt ggctgttaac taaaaggttg gtggttcgaa 60

cccacccaga ggcg 74

<210> 60

<211> 74

<212> DNA

<213> Intelligent people

<400> 60

gtctctgtgg cgcaatcggt tagcgcgttc ggctgttaac tgaaaggtta gtggttcgag 60

cccacccggg gacg 74

<210> 61

<211> 72

<212> DNA

<213> Intelligent people

<400> 61

tcctcgttag tatagtggtt agtatccccg cctgtcacgc gggagaccgg ggttcaattc 60

cccgacgggg ag 72

<210> 62

<211> 72

<212> DNA

<213> Intelligent people

<400> 62

tcctcgttag tatagtggtg agtatccccg cctgtcacgc gggagaccgg ggttcgattc 60

cccgacgggg ag 72

<210> 63

<211> 72

<212> DNA

<213> Intelligent people

<400> 63

tcctcgttag tatagtggtg agtgtccccg tctgtcacgc gggagaccgg ggttcgattc 60

cccgacgggg ag 72

<210> 64

<211> 72

<212> DNA

<213> Intelligent people

<400> 64

gggggcatag ctcagtggta gagcatttga ctgcagatca agaggtccct ggttcaaatc 60

caggtgcccc ct 72

<210> 65

<211> 72

<212> DNA

<213> Intelligent people

<400> 65

gggggtatag ctcaggggta gagcatttga ctgcagatca agaggtccct ggttcaaatc 60

caggtgcccc cc 72

<210> 66

<211> 72

<212> DNA

<213> Intelligent people

<400> 66

gggggtatag cttagcggta gagcatttga ctgcagatca agaggtcccc ggttcaaatc 60

cgggtgcccc ct 72

<210> 67

<211> 72

<212> DNA

<213> Intelligent people

<400> 67

gggggtatag cttaggggta gagcatttga ctgcagatca aaaggtccct ggttcaaatc 60

caggtgcccc tt 72

<210> 68

<211> 72

<212> DNA

<213> Intelligent people

<400> 68

gggggtatag ctcaggggta gagcatttga ctgcagatca agaggtcccc agttcaaatc 60

tgggtgcccc ct 72

<210> 69

<211> 72

<212> DNA

<213> Intelligent people

<400> 69

gggggtatag ctcaggggta gagcatttga ctgcagatca agaagtcccc ggttcaaatc 60

cgggtgcccc ct 72

<210> 70

<211> 72

<212> DNA

<213> Intelligent people

<400> 70

gggggtatag ctcaggggta gagcatttga ctgcagatca agaggtctct ggttcaaatc 60

caggtgcccc ct 72

<210> 71

<211> 72

<212> DNA

<213> Intelligent people

<400> 71

gggggtatag ctcaggggta gagcacttga ctgcagatca agaagtcctt ggttcaaatc 60

caggtgcccc ct 72

<210> 72

<211> 72

<212> DNA

<213> Intelligent people

<400> 72

ggggatatag ctcaggggta gagcatttga ctgcagatca agaggtcccc ggttcaaatc 60

cgggtgcccc cc 72

<210> 73

<211> 72

<212> DNA

<213> Intelligent people

<400> 73

gggggtatag ttcaggggta gagcatttga ctgcagatca agaggtccct ggttcaaatc 60

caggtgcccc ct 72

<210> 74

<211> 72

<212> DNA

<213> Intelligent people

<400> 74

gggggtatag ctcaggggta gagcatttga ctgcaaatca agaggtccct gattcaaatc 60

caggtgcccc ct 72

<210> 75

<211> 72

<212> DNA

<213> Intelligent people

<400> 75

gggggtatag ctcagtggta gagcatttga ctgcagatca agaggtcccc ggttcaaatc 60

cgggtgcccc ct 72

<210> 76

<211> 72

<212> DNA

<213> Intelligent people

<400> 76

gggcgtatag ctcaggggta gagcatttga ctgcagatca agaggtcccc agttcaaatc 60

tgggtgcccc ct 72

<210> 77

<211> 72

<212> DNA

<213> Intelligent people

<400> 77

gggggtatag ctcacaggta gagcatttga ctgcagatca agaggtcccc ggttcaaatc 60

tgggtgcccc ct 72

<210> 78

<211> 70

<212> DNA

<213> Intelligent people

<400> 78

gggcgtatag ctcaggggta gagcatttga ctgcagatca agaggtcccc agttcaaatc 60

tgggtgccca 70

<210> 79

<211> 72

<212> DNA

<213> Intelligent people

<400> 79

gggggtatag ctcacaggta gagcatttga ctgcagatca agaggtcccc ggttcaaatc 60

cggttactcc ct 72

<210> 80

<211> 72

<212> DNA

<213> Intelligent people

<400> 80

gggggtatag ctcaggggta gagcacttga ctgcagatca agaggtccct ggttcaaatc 60

caggtgcccc ct 72

<210> 81

<211> 72

<212> DNA

<213> Intelligent people

<400> 81

gggggtatag ctcagtggta gagcatttga ctgcagatca agaggtccct ggttcaaatc 60

cgggtgcccc ct 72

<210> 82

<211> 73

<212> DNA

<213> Intelligent people

<400> 82

gggggtatag ctcagtgggt agagcatttg actgcagatc aagaggtccc cggttcaaat 60

ccgggtgccc cct 73

<210> 83

<211> 72

<212> DNA

<213> Intelligent people

<400> 83

gggggtgtag ctcagtggta gagcatttga ctgcagatca agaggtccct ggttcaaatc 60

caggtgcccc ct 72

<210> 84

<211> 73

<212> DNA

<213> Intelligent people

<400> 84

gggggtatag ctcaggtggt agagcatttg actgcagatc aagaggtccc cggttcaaat 60

ccgggtgccc cct 73

<210> 85

<211> 72

<212> DNA

<213> Intelligent people

<400> 85

gggggtatag ctcaggggta gagcatttga ctgcagatca agaggtcccc ggttcaaatc 60

cgggtgcccc ct 72

<210> 86

<211> 72

<212> DNA

<213> Intelligent people

<400> 86

gggggtatag ctcaggggta gagcatttga ctgcagatca agaggtccct ggttcaaatc 60

caggtgcccc ct 72

<210> 87

<211> 72

<212> DNA

<213> Intelligent people

<400> 87

ggttccatgg tgtaatggtt agcactctgg actctgaatc cagcgatccg agttcaaatc 60

tcggtggaac ct 72

<210> 88

<211> 72

<212> DNA

<213> Intelligent people

<400> 88

ggttccatgg tgtaatggtt agcactctgg actctgaatc cagcgatccg agttcaagtc 60

tcggtggaac ct 72

<210> 89

<211> 72

<212> DNA

<213> Intelligent people

<400> 89

ggttccatgg tgtaatggtg agcactctgg actctgaatc cagcgatccg agttcgagtc 60

tcggtggaac ct 72

<210> 90

<211> 72

<212> DNA

<213> Intelligent people

<400> 90

ggttccatgg tgtaatggta agcactctgg actctgaatc cagcgatccg agttcgagtc 60

tcggtggaac ct 72

<210> 91

<211> 72

<212> DNA

<213> Intelligent people

<400> 91

ggttccatgg tgtaatggtt agcactctgg actctgaatc cggtaatccg agttcaaatc 60

tcggtggaac ct 72

<210> 92

<211> 72

<212> DNA

<213> Intelligent people

<400> 92

ggccccatgg tgtaatggtc agcactctgg actctgaatc cagcgatccg agttcaaatc 60

tcggtgggac cc 72

<210> 93

<211> 72

<212> DNA

<213> Intelligent people

<400> 93

ggttccatgg tgtaatggta agcactctgg actctgaatc cagccatctg agttcgagtc 60

tctgtggaac ct 72

<210> 94

<211> 72

<212> DNA

<213> Intelligent people

<400> 94

ggtcccatgg tgtaatggtt agcactctgg actttgaatc cagcgatccg agttcaaatc 60

tcggtgggac ct 72

<210> 95

<211> 72

<212> DNA

<213> Intelligent people

<400> 95

ggtcccatgg tgtaatggtt agcactctgg actttgaatc cagcaatccg agttcgaatc 60

tcggtgggac ct 72

<210> 96

<211> 72

<212> DNA

<213> Intelligent people

<400> 96

ggccccatgg tgtaatggtt agcactctgg actttgaatc cagcgatccg agttcaaatc 60

tcggtgggac ct 72

<210> 97

<211> 72

<212> DNA

<213> Intelligent people

<400> 97

ggtcccatgg tgtaatggtt agcactctgg gctttgaatc cagcaatccg agttcgaatc 60

ttggtgggac ct 72

<210> 98

<211> 72

<212> DNA

<213> Intelligent people

<400> 98

tccctggtgg tctagtggtt aggattcggc gctctcaccg ccgcggcccg ggttcgattc 60

ccggtcaggg aa 72

<210> 99

<211> 72

<212> DNA

<213> Intelligent people

<400> 99

tccctggtgg tctagtggtt aggattcggc gctctcaccg ccgcggcccg ggttcgattc 60

ccggtcagga aa 72

<210> 100

<211> 72

<212> DNA

<213> Intelligent people

<400> 100

tcccatatgg tctagcggtt aggattcctg gttttcaccc aggtggcccg ggttcgactc 60

ccggtatggg aa 72

<210> 101

<211> 72

<212> DNA

<213> Intelligent people

<400> 101

tcccacatgg tctagcggtt aggattcctg gttttcaccc aggcggcccg ggttcgactc 60

ccggtgtggg aa 72

<210> 102

<211> 72

<212> DNA

<213> Intelligent people

<400> 102

tccctggtgg tctagtggct aggattcggc gctttcaccg ccgcggcccg ggttcgattc 60

ccggccaggg aa 72

<210> 103

<211> 72

<212> DNA

<213> Intelligent people

<400> 103

tccctggtgg tctagtggct aggattcggc gctttcaccg ccgcggcccg ggttcgattc 60

ccggtcaggg aa 72

<210> 104

<211> 71

<212> DNA

<213> Intelligent people

<400> 104

gcattggtgg ttcagtggta gaattctcgc ctcccacgcg ggagacccgg gttcaattcc 60

cggccaatgc a 71

<210> 105

<211> 71

<212> DNA

<213> Intelligent people

<400> 105

gcgccgctgg tgtagtggta tcatgcaaga ttcccattct tgcgacccgg gttcgattcc 60

cgggcggcgc a 71

<210> 106

<211> 71

<212> DNA

<213> Intelligent people

<400> 106

gcattggtgg ttcaatggta gaattctcgc ctcccacgca ggagacccag gttcgattcc 60

tggccaatgc a 71

<210> 107

<211> 71

<212> DNA

<213> Intelligent people

<400> 107

gcatgggtgg ttcagtggta gaattctcgc ctgccacgcg ggaggcccgg gttcgattcc 60

cggcccatgc a 71

<210> 108

<211> 71

<212> DNA

<213> Intelligent people

<400> 108

gcattggtgg ttcagtggta gaattctcgc ctgccacgcg ggaggcccgg gttcgattcc 60

cggccaatgc a 71

<210> 109

<211> 71

<212> DNA

<213> Intelligent people

<400> 109

gcattggtgg ttcagtggta gaattctcgc ctgccacgcg ggaggcccgg gtttgattcc 60

cggccagtgc a 71

<210> 110

<211> 71

<212> DNA

<213> Intelligent people

<400> 110

gcataggtgg ttcagtggta gaattcttgc ctgccacgca ggaggcccag gtttgattcc 60

tggcccatgc a 71

<210> 111

<211> 71

<212> DNA

<213> Intelligent people

<400> 111

gcattggtgg ttcagtggta gaattctcgc ctgccatgcg ggcggccggg cttcgattcc 60

tggccaatgc a 71

<210> 112

<211> 72

<212> DNA

<213> Intelligent people

<400> 112

gcgttggtgg tatagtggtt agcatagctg ccttccaagc agttgacccg ggttcgattc 60

ccggccaacg ca 72

<210> 113

<211> 72

<212> DNA

<213> Intelligent people

<400> 113

gcgttggtgg tatagtggtg agcatagctg ccttccaagc agttgacccg ggttcgattc 60

ccggccaacg ca 72

<210> 114

<211> 72

<212> DNA

<213> Intelligent people

<400> 114

gcgttggtgg tatagtggta agcatagctg ccttccaagc agttgacccg ggttcgattc 60

ccggccaacg ca 72

<210> 115

<211> 72

<212> DNA

<213> Intelligent people

<400> 115

gcgttggtgg tatagtggtg agcatagttg ccttccaagc agttgacccg ggctcgattc 60

ccgcccaacg ca 72

<210> 116

<211> 72

<212> DNA

<213> Intelligent people

<400> 116

gccgtgatcg tatagtggtt agtactctgc gttgtggccg cagcaacctc ggttcgaatc 60

cgagtcacgg ca 72

<210> 117

<211> 72

<212> DNA

<213> Intelligent people

<400> 117

gccatgatcg tatagtggtt agtactctgc gctgtggccg cagcaacctc ggttcgaatc 60

cgagtcacgg ca 72

<210> 118

<211> 74

<212> DNA

<213> Intelligent people

<400> 118

ggccggttag ctcagttggt tagagcgtgg cgctaataac gccaaggtcg cgggttcgat 60

ccccgtacgg gcca 74

<210> 119

<211> 74

<212> DNA

<213> Intelligent people

<400> 119

ggccggttag ctcagttggt tagagcgtgg tgctaataac gccaaggtcg cgggttcgat 60

ccccgtactg gcca 74

<210> 120

<211> 74

<212> DNA

<213> Intelligent people

<400> 120

ggctggttag ctcagttggt tagagcgtgg tgctaataac gccaaggtcg cgggttcgat 60

ccccgtactg gcca 74

<210> 121

<211> 74

<212> DNA

<213> Intelligent people

<400> 121

ggccggttag ctcagttggt tagagcgtgg tgctaataac gccaaggtcg cgggttcgaa 60

ccccgtacgg gcca 74

<210> 122

<211> 74

<212> DNA

<213> Intelligent people

<400> 122

ggccggttag ctcagttggt tagagcgtgg tgctaataac gccaaggtcg cgggttcgat 60

ccccgtacgg gcca 74

<210> 123

<211> 74

<212> DNA

<213> Intelligent people

<400> 123

ggccggttag ctcagttggt tagagcgtgg tgctaataac gctaaggtcg cgggttcgat 60

ccccgtactg gcca 74

<210> 124

<211> 74

<212> DNA

<213> Intelligent people

<400> 124

ggccggttag ctcagttggt cagagcgtgg tgctaataac gccaaggtcg cgggttcgat 60

ccccgtacgg gcca 74

<210> 125

<211> 74

<212> DNA

<213> Intelligent people

<400> 125

ggccggttag ctcagtcggc tagagcgtgg tgctaataac gccaaggtcg cgggttcgat 60

ccccgtacgg gcca 74

<210> 126

<211> 74

<212> DNA

<213> Intelligent people

<400> 126

ggctggttag ttcagttggt tagagcgtgg tgctaataac gccaaggtcg tgggttcgat 60

ccccatatcg gcca 74

<210> 127

<211> 74

<212> DNA

<213> Intelligent people

<400> 127

ggccggttag ctcagttggt aagagcgtgg tgctgataac accaaggtcg cgggctcgac 60

tcccgcaccg gcca 74

<210> 128

<211> 93

<212> DNA

<213> Intelligent people

<400> 128

gctccagtgg cgcaatcggt tagcgcgcgg tacttatatg acagtgcgag cggagcaatg 60

ccgaggttgt gagttcgatc ctcacctgga gca 93

<210> 129

<211> 93

<212> DNA

<213> Intelligent people

<400> 129

gctccagtgg cgcaatcggt tagcgcgcgg tacttataca gcagtacatg cagagcaatg 60

ccgaggttgt gagttcgagc ctcacctgga gca 93

<210> 130

<211> 94

<212> DNA

<213> Intelligent people

<400> 130

gctccagtgg cgcaatcggt tagcgcgcgg tacttatatg gcagtatgtg tgcgagtgat 60

gccgaggttg tgagttcgag cctcacctgg agca 94

<210> 131

<211> 94

<212> DNA

<213> Intelligent people

<400> 131

gctccagtgg cgcaatcggt tagcgcgcgg tacttataca acagtatatg tgcgggtgat 60

gccgaggttg tgagttcgag cctcacctgg agca 94

<210> 132

<211> 94

<212> DNA

<213> Intelligent people

<400> 132

gctccagtgg cgcaatcggt tagcgcgcgg tacttataag acagtgcacc tgtgagcaat 60

gccgaggttg tgagttcaag cctcacctgg agca 94

<210> 133

<211> 82

<212> DNA

<213> Intelligent people

<400> 133

ggtagcgtgg ccgagcggtc taaggcgctg gattaaggct ccagtctctt cggaggcgtg 60

ggttcgaatc ccaccgctgc ca 82

<210> 134

<211> 82

<212> DNA

<213> Intelligent people

<400> 134

ggtagcgtgg ccgagcggtc taaggcgctg gattaaggct ccagtctctt cgggggcgtg 60

ggttcgaatc ccaccgctgc ca 82

<210> 135

<211> 82

<212> DNA

<213> Intelligent people

<400> 135

ggtagcgtgg ccgagcggtc taaggcgctg gattaaggct ccagtctctt cgggggcgtg 60

ggttcaaatc ccaccgctgc ca 82

<210> 136

<211> 82

<212> DNA

<213> Intelligent people

<400> 136

ggtagcgtgg ccgagtggtc taagacgctg gattaaggct ccagtctctt cgggggcgtg 60

ggtttgaatc ccaccgctgc ca 82

<210> 137

<211> 106

<212> DNA

<213> Intelligent people

<400> 137

gtcaggatgg ccgagtggtc taaggcgcca gactcaagct aagcttcctc cgcggtgggg 60

attctggtct ccaatggagg cgtgggttcg aatcccactt ctgaca 106

<210> 138

<211> 105

<212> DNA

<213> Intelligent people

<400> 138

gtcaggatgg ccgagtggtc taaggcgcca gactcaagct tggcttcctc gtgttgagga 60

ttctggtctc caatggaggc gtgggttcga atcccacttc tgaca 105

<210> 139

<211> 108

<212> DNA

<213> Intelligent people

<400> 139

gtcaggatgg ccgagtggtc taaggcgcca gactcaagct tactgcttcc tgtgttcggg 60

tcttctggtc tccgtatgga ggcgtgggtt cgaatcccac ttctgaca 108

<210> 140

<211> 107

<212> DNA

<213> Intelligent people

<400> 140

gtcaggatgg ccgagtggtc taaggcgcca gactcaagtt gctacttccc aggtttgggg 60

cttctggtct ccgcatggag gcgtgggttc gaatcccact tctgaca 107

<210> 141

<211> 106

<212> DNA

<213> Intelligent people

<400> 141

gtcaggatgg ccgagtggtc taaggcgcca gactcaaggt aagcaccttg cctgcgggct 60

ttctggtctc cggatggagg cgtgggttcg aatcccactt ctgaca 106

<210> 142

<211> 74

<212> DNA

<213> Intelligent people

<400> 142

gcctccttag tgcagtaggt agcgcatcag tctcaaaatc tgaatggtcc tgagttcaag 60

cctcagaggg ggca 74

<210> 143

<211> 84

<212> DNA

<213> Intelligent people

<400> 143

gtcaggatgg ccgagcagtc ttaaggcgct gcgttcaaat cgcaccctcc gctggaggcg 60

tgggttcgaa tcccactttt gaca 84

<210> 144

<211> 83

<212> DNA

<213> Intelligent people

<400> 144

gtcaggatgg ccgagcggtc taaggcgctg cgttcaggtc gcagtctccc ctggaggcgt 60

gggttcgaat cccactcctg aca 83

<210> 145

<211> 83

<212> DNA

<213> Intelligent people

<400> 145

gtcaggatgg ccgagcggtc taaggcgctg cgttcaggtc gcagtctccc ctggaggcgt 60

gggttcgaat cccacttctg aca 83

<210> 146

<211> 83

<212> DNA

<213> Intelligent people

<400> 146

accaggatgg ccgagtggtt aaggcgttgg acttaagatc caatggacat atgtccgcgt 60

gggttcgaac cccactcctg gta 83

<210> 147

<211> 83

<212> DNA

<213> Intelligent people

<400> 147

accgggatgg ccgagtggtt aaggcgttgg acttaagatc caatgggctg gtgcccgcgt 60

gggttcgaac cccactctcg gta 83

<210> 148

<211> 83

<212> DNA

<213> Intelligent people

<400> 148

accagaatgg ccgagtggtt aaggcgttgg acttaagatc caatggattc atatccgcgt 60

gggttcgaac cccacttctg gta 83

<210> 149

<211> 83

<212> DNA

<213> Intelligent people

<400> 149

accgggatgg ctgagtggtt aaggcgttgg acttaagatc caatggacag gtgtccgcgt 60

gggttcgagc cccactcccg gta 83

<210> 150

<211> 82

<212> DNA

<213> Intelligent people

<400> 150

ggtagcgtgg ccgagcggtc taaggcgctg gatttaggct ccagtctctt cggaggcgtg 60

ggttcgaatc ccaccgctgc ca 82

<210> 151

<211> 82

<212> DNA

<213> Intelligent people

<400> 151

ggtagtgtgg ccgagcggtc taaggcgctg gatttaggct ccagtctctt cgggggcgtg 60

ggttcgaatc ccaccactgc ca 82

<210> 152

<211> 82

<212> DNA

<213> Intelligent people

<400> 152

ggtagcgtgg ccgagtggtc taaggcgctg gatttaggct ccagtcattt cgatggcgtg 60

ggttcgaatc ccaccgctgc ca 82

<210> 153

<211> 73

<212> DNA

<213> Intelligent people

<400> 153

gcccggctag ctcagtcggt agagcatggg actcttaatc ccagggtcgt gggttcgagc 60

cccacgttgg gcg 73

<210> 154

<211> 73

<212> DNA

<213> Intelligent people

<400> 154

gcccagctag ctcagtcggt agagcataag actcttaatc tcagggttgt ggattcgtgc 60

cccatgctgg gtg 73

<210> 155

<211> 74

<212> DNA

<213> Intelligent people

<400> 155

gcagctagct cagtcggtag agcatgagac tcttaatctc agggtcatgg gttcgtgccc 60

catgttgggt gcca 74

<210> 156

<211> 73

<212> DNA

<213> Intelligent people

<400> 156

gcccggctag ctcagtcggt agagcatgag actcttaatc tcagggtcgt gggttcgagc 60

cccacgttgg gcg 73

<210> 157

<211> 73

<212> DNA

<213> Intelligent people

<400> 157

gcccggctag ctcagtcggt agagcatgag acccttaatc tcagggtcgt gggttcgagc 60

cccacgttgg gcg 73

<210> 158

<211> 73

<212> DNA

<213> Intelligent people

<400> 158

gcccggctag ctcagtcggt agagcatggg actcttaatc tcagggtcgt gggttcgagc 60

cccacgttgg gcg 73

<210> 159

<211> 73

<212> DNA

<213> Intelligent people

<400> 159

gcccggctag ctcagtcgat agagcatgag actcttaatc tcagggtcgt gggttcgagc 60

cgcacgttgg gcg 73

<210> 160

<211> 73

<212> DNA

<213> Intelligent people

<400> 160

gcccagctag ctcagtcggt agagcatgag actcttaatc tcagggtcat gggtttgagc 60

cccacgtttg gtg 73

<210> 161

<211> 73

<212> DNA

<213> Intelligent people

<400> 161

gcctggctag ctcagtcggc aaagcatgag actcttaatc tcagggtcgt gggctcgagc 60

tccatgttgg gcg 73

<210> 162

<211> 73

<212> DNA

<213> Intelligent people

<400> 162

gcccgactac ctcagtcggt ggagcatggg actcttcatc ccagggttgt gggttcgagc 60

cccacattgg gca 73

<210> 163

<211> 73

<212> DNA

<213> Intelligent people

<400> 163

gcctggatag ctcagttggt agagcatcag acttttaatc tgagggtcca gggttcaagt 60

ccctgttcag gca 73

<210> 164

<211> 73

<212> DNA

<213> Intelligent people

<400> 164

acccagatag ctcagtcagt agagcatcag acttttaatc tgagggtcca aggttcatgt 60

ccctttttgg gtg 73

<210> 165

<211> 73

<212> DNA

<213> Intelligent people

<400> 165

gcctggatag ctcagttggt agagcatcag acttttaatc tgagggtcca gggttcaagt 60

ccctgttcag gcg 73

<210> 166

<211> 73

<212> DNA

<213> Intelligent people

<400> 166

gcccggatag ctcagtcggt agagcatcag acttttaatc tgagggtcca gggttcaagt 60

ccctgttcgg gcg 73

<210> 167

<211> 73

<212> DNA

<213> Intelligent people

<400> 167

gcctggatag ctcagtcggt agagcatcag acttttaatc tgagggtcca gggttcaagt 60

ccctgttcag gcg 73

<210> 168

<211> 73

<212> DNA

<213> Intelligent people

<400> 168

gcccggatag ctcagtcggt agagcatcag acttttaatc tgagggtccg gggttcaagt 60

ccctgttcgg gcg 73

<210> 169

<211> 73

<212> DNA

<213> Intelligent people

<400> 169

gcctgggtag ctcagtcggt agagcatcag acttttaatc tgagggtcca gggttcaagt 60

ccctgtccag gcg 73

<210> 170

<211> 73

<212> DNA

<213> Intelligent people

<400> 170

gcctggatag ctcagttggt agaacatcag acttttaatc tgacggtgca gggttcaagt 60

ccctgttcag gcg 73

<210> 171

<211> 73

<212> DNA

<213> Intelligent people

<400> 171

gcctcgttag cgcagtaggt agcgcgtcag tctcataatc tgaaggtcgt gagttcgatc 60

ctcacacggg gca 73

<210> 172

<211> 73

<212> DNA

<213> Intelligent people

<400> 172

gccctcttag cgcagtgggc agcgcgtcag tctcataatc tgaaggtcct gagttcgagc 60

ctcagagagg gca 73

<210> 173

<211> 73

<212> DNA

<213> Intelligent people

<400> 173

gcctccttag cgcagtaggc agcgcgtcag tctcataatc tgaaggtcct gagttcgaac 60

ctcagagggg gca 73

<210> 174

<211> 73

<212> DNA

<213> Intelligent people

<400> 174

gccctcttag cgcagcgggc agcgcgtcag tctcataatc tgaaggtcct gagttcgagc 60

ctcagagagg gca 73

<210> 175

<211> 73

<212> DNA

<213> Intelligent people

<400> 175

gccctcttag cgcagctggc agcgcgtcag tctcataatc tgaaggtcct gagttcaagc 60

ctcagagagg gca 73

<210> 176

<211> 73

<212> DNA

<213> Intelligent people

<400> 176

gcctcgttag cgcagtaggc agcgcgtcag tctcataatc tgaaggtcgt gagttcgagc 60

ctcacacggg gca 73

<210> 177

<211> 73

<212> DNA

<213> Intelligent people

<400> 177

gccctcttag tgcagctggc agcgcgtcag tttcataatc tgaaagtcct gagttcaagc 60

ctcagagagg gca 73

<210> 178

<211> 73

<212> DNA

<213> Intelligent people

<400> 178

gccgaaatag ctcagttggg agagcgttag actgaagatc taaaggtccc tggttcgatc 60

ccgggtttcg gca 73

<210> 179

<211> 73

<212> DNA

<213> Intelligent people

<400> 179

gccgaaatag ctcagttggg agagcgttag actgaagatc taaaggtccc tggttcaatc 60

ccgggtttcg gca 73

<210> 180

<211> 73

<212> DNA

<213> Intelligent people

<400> 180

gccgagatag ctcagttggg agagcgttag actgaagatc taaaggtccc tggttcaatc 60

ccgggtttcg gca 73

<210> 181

<211> 74

<212> DNA

<213> Intelligent people

<400> 181

gccgaaatag ctcagttggg agagcgttag accgaagatc ttaaaggtcc ctggttcaat 60

cccgggtttc ggca 74

<210> 182

<211> 74

<212> DNA

<213> Intelligent people

<400> 182

gctgaaatag ctcagttggg agagcgttag actgaagatc ttaaagttcc ctggttcaac 60

cctgggtttc agcc 74

<210> 183

<211> 72

<212> DNA

<213> Intelligent people

<400> 183

ggctcgttgg tctaggggta tgattctcgc ttaggatgcg agaggtcccg ggttcaaatc 60

ccggacgagc cc 72

<210> 184

<211> 72

<212> DNA

<213> Intelligent people

<400> 184

ggctcgttgg tctaggggta tgattctcgc ttagggtgcg agaggtcccg ggttcaaatc 60

ccggacgagc cc 72

<210> 185

<211> 72

<212> DNA

<213> Intelligent people

<400> 185

ggctcgttgg tctaggggta tgattctcgc ttcgggtgcg agaggtcccg ggttcaaatc 60

ccggacgagc cc 72

<210> 186

<211> 72

<212> DNA

<213> Intelligent people

<400> 186

ggctcgttgg tctaggggta tgattctcgc ttcgggtgtg agaggtcccg ggttcaaatc 60

ccggacgagc cc 72

<210> 187

<211> 72

<212> DNA

<213> Intelligent people

<400> 187

ggctcgttgg tctagtggta tgattctcgc tttgggtgcg agaggtcccg ggttcaaatc 60

ccggacgagc cc 72

<210> 188

<211> 72

<212> DNA

<213> Intelligent people

<400> 188

ggctcgttgg tctaggggta tgattctcgg tttgggtccg agaggtcccg ggttcaaatc 60

ccggacgagc cc 72

<210> 189

<211> 72

<212> DNA

<213> Intelligent people

<400> 189

ggctcgttgg tctaggggta tgattctcgc tttgggtgcg agaggtcccg ggttcaaatc 60

ccggacgagc cc 72

<210> 190

<211> 87

<212> DNA

<213> Intelligent people

<400> 190

gcccggatga tcctcagtgg tctggggtgc aggcttcaaa cctgtagctg tctagcgaca 60

gagtggttca attccacctt tcgggcg 87

<210> 191

<211> 84

<212> DNA

<213> Intelligent people

<400> 191

gctcggatga tcctcagtgg tctggggtgc aggcttcaaa cctgtagctg tctagtgaca 60

gagtggttca attccacctt tgta 84

<210> 192

<211> 82

<212> DNA

<213> Intelligent people

<400> 192

gtagtcgtgg ccgagtggtt aaggcgatgg actagaaatc cattggggtt tccccgcgca 60

ggttcgaatc ctgccgacta cg 82

<210> 193

<211> 82

<212> DNA

<213> Intelligent people

<400> 193

gtagtcgtgg ccgagtggtt aaggcgatgg actagaaatc cattggggtc tccccgcgca 60

ggttcgaatc ctgccgacta cg 82

<210> 194

<211> 82

<212> DNA

<213> Intelligent people

<400> 194

gtagtcgtgg ccgagtggtt aaggcgatgg actagaaatc cattggggtt tccccacgca 60

ggttcgaatc ctgccgacta cg 82

<210> 195

<211> 82

<212> DNA

<213> Intelligent people

<400> 195

gtagtcgtgg ccgagtggtt aaggtgatgg actagaaacc cattggggtc tccccgcgca 60

ggttcgaatc ctgccgacta cg 82

<210> 196

<211> 82

<212> DNA

<213> Intelligent people

<400> 196

gctgtgatgg ccgagtggtt aaggcgttgg actcgaaatc caatggggtc tccccgcgca 60

ggttcgaatc ctgctcacag cg 82

<210> 197

<211> 82

<212> DNA

<213> Intelligent people

<400> 197

gctgtgatgg ccgagtggtt aaggcgttgg actcgaaatc caatggggtc tccccgcgca 60

ggttcaaatc ctgctcacag cg 82

<210> 198

<211> 82

<212> DNA

<213> Intelligent people

<400> 198

gctgtgatgg ccgagtggtt aaggtgttgg actcgaaatc caatgggggt tccccgcgca 60

ggttcaaatc ctgctcacag cg 82

<210> 199

<211> 82

<212> DNA

<213> Intelligent people

<400> 199

gtcacggtgg ccgagtggtt aaggcgttgg actcgaaatc caatggggtt tccccgcaca 60

ggttcgaatc ctgttcgtga cg 82

<210> 200

<211> 82

<212> DNA

<213> Intelligent people

<400> 200

gacgaggtgg ccgagtggtt aaggcgatgg actgctaatc cattgtgctc tgcacgcgtg 60

ggttcgaatc ccaccctcgt cg 82

<210> 201

<211> 82

<212> DNA

<213> Intelligent people

<400> 201

gacgaggtgg ccgagtggtt aaggcgatgg actgctaatc cattgtgctc tgcacgcgtg 60

ggttcgaatc ccaccttcgt cg 82

<210> 202

<211> 82

<212> DNA

<213> Intelligent people

<400> 202

gacgaggtgg ccgagtggtt aaggcgatgg actgctaatc cattgtgctt tgcacgcgtg 60

ggttcgaatc ccatcctcgt cg 82

<210> 203

<211> 82

<212> DNA

<213> Intelligent people

<400> 203

gacgaggtgg ccgagtggtt aaggcgatgg actgctaatc cattgtgctc tgcacgcgtg 60

ggttcgaatc ccatcctcgt cg 82

<210> 204

<211> 82

<212> DNA

<213> Intelligent people

<400> 204

gacgaggtgg ccgagtggtt aaggcgatgg actgctaatc cattgtgctc tgcacacgtg 60

ggttcgaatc ccatcctcgt cg 82

<210> 205

<211> 84

<212> DNA

<213> Intelligent people

<400> 205

ggagaggcct ggccgagtgg ttaaggcgat ggactgctaa tccattgtgc tctgcacgcg 60

tgggttcgaa tcccatcctc gtcg 84

<210> 206

<211> 82

<212> DNA

<213> Intelligent people

<400> 206

gcagcgatgg ccgagtggtt aaggcgttgg acttgaaatc caatggggtc tccccgcgca 60

ggttcgaacc ctgctcgctg cg 82

<210> 207

<211> 82

<212> DNA

<213> Intelligent people

<400> 207

gtagtcgtgg ccgagtggtt aaggcgatgg acttgaaatc cattggggtt tccccgcgca 60

ggttcgaatc ctgccgacta cg 82

<210> 208

<211> 82

<212> DNA

<213> Intelligent people

<400> 208

gtagtcgtgg ccgagtggtt aaggcgatgg acttgaaatc cattggggtc tccccgcgca 60

ggttcgaatc ctgccgacta cg 82

<210> 209

<211> 82

<212> DNA

<213> Intelligent people

<400> 209

gtagtcgtgg ccgagtggtt aaggcgatgg acttgaaatc cattggggtt tccccgcgca 60

ggttcgaatc ctgtcggcta cg 82

<210> 210

<211> 74

<212> DNA

<213> Intelligent people

<400> 210

ggcgccgtgg cttagttggt taaagcgcct gtctagtaaa caggagatcc tgggttcgaa 60

tcccagcggt gcct 74

<210> 211

<211> 74

<212> DNA

<213> Intelligent people

<400> 211

ggctccgtgg cttagctggt taaagcgcct gtctagtaaa caggagatcc tgggttcgaa 60

tcccagcggg gcct 74

<210> 212

<211> 74

<212> DNA

<213> Intelligent people

<400> 212

ggctccgtag cttagttggt taaagcgcct gtctagtaaa caggagatcc tgggttcgac 60

tcccagcggg gcct 74

<210> 213

<211> 74

<212> DNA

<213> Intelligent people

<400> 213

ggcttcgtgg cttagctggt taaagcgcct gtctagtaaa caggagatcc tgggttcgaa 60

tcccagcgag gcct 74

<210> 214

<211> 74

<212> DNA

<213> Intelligent people

<400> 214

ggcgccgtgg cttagctggt taaagcgcct gtctagtaaa caggagatcc tgggttcgaa 60

tcccagcggt gcct 74

<210> 215

<211> 74

<212> DNA

<213> Intelligent people

<400> 215

ggccctgtgg cttagctggt caaagcgcct gtctagtaaa caggagatcc tgggttcgaa 60

tcccagcggg gcct 74

<210> 216

<211> 74

<212> DNA

<213> Intelligent people

<400> 216

ggctctatgg cttagttggt taaagcgcct gtctcgtaaa caggagatcc tgggttcgac 60

tcccagtggg gcct 74

<210> 217

<211> 72

<212> DNA

<213> Intelligent people

<400> 217

ggcgcggtgg ccaagtggta aggcgtcggt ctcgtaaacc gaagatcacg ggttcgaacc 60

ccgtccgtgc ct 72

<210> 218

<211> 74

<212> DNA

<213> Intelligent people

<400> 218

ggctctgtgg cttagttggc taaagcgcct gtctcgtaaa caggagatcc tgggttcgaa 60

tcccagcggg gcct 74

<210> 219

<211> 72

<212> DNA

<213> Intelligent people

<400> 219

ggcgcggtgg ccaagtggta aggcgtcggt ctcgtaaacc gaagatcgcg ggttcgaacc 60

ccgtccgtgc ct 72

<210> 220

<211> 74

<212> DNA

<213> Intelligent people

<400> 220

ggccctgtag ctcagcggtt ggagcgctgg tctcgtaaac ctaggggtcg tgagttcaaa 60

tctcaccagg gcct 74

<210> 221

<211> 74

<212> DNA

<213> Intelligent people

<400> 221

ggctctatgg cttagttggt taaagcgcct gtcttgtaaa caggagatcc tgggttcgaa 60

tcccagtaga gcct 74

<210> 222

<211> 73

<212> DNA

<213> Intelligent people

<400> 222

ggctccatag ctcagtggtt agagcactgg tcttgtaaac caggggtcgc gagttcgatc 60

ctcgctgggg cct 73

<210> 223

<211> 73

<212> DNA

<213> Intelligent people

<400> 223

ggctccatag ctcaggggtt agagcgctgg tcttgtaaac caggggtcgc gagttcaatt 60

ctcgctgggg cct 73

<210> 224

<211> 73

<212> DNA

<213> Intelligent people

<400> 224

ggctccatag ctcaggggtt agagcactgg tcttgtaaac caggggtcgc gagttcaaat 60

ctcgctgggg cct 73

<210> 225

<211> 73

<212> DNA

<213> Intelligent people

<400> 225

ggccctatag ctcaggggtt agagcactgg tcttgtaaac caggggtcgc gagttcaaat 60

ctcgctgggg cct 73

<210> 226

<211> 72

<212> DNA

<213> Intelligent people

<400> 226

ggctccatag ctcaggggtt agagcactgg tcttgtaaac cagggtcgcg agttcaaatc 60

tcgctggggc ct 72

<210> 227

<211> 72

<212> DNA

<213> Intelligent people

<400> 227

ggcctcgtgg cgcaacggta gcgcgtctga ctccagatca gaaggttgcg tgttcaaatc 60

acgtcggggt ca 72

<210> 228

<211> 72

<212> DNA

<213> Intelligent people

<400> 228

gacctcgtgg cgcaatggta gcgcgtctga ctccagatca gaaggttgcg tgttcaagtc 60

acgtcggggt ca 72

<210> 229

<211> 72

<212> DNA

<213> Intelligent people

<400> 229

gacctcgtgg cgcaacggta gcgcgtctga ctccagatca gaaggttgcg tgttcaaatc 60

acgtcggggt ca 72

<210> 230

<211> 72

<212> DNA

<213> Intelligent people

<400> 230

gacctcgtgg cgcaacggta gcgcgtctga ctccagatca gaaggctgcg tgttcgaatc 60

acgtcggggt ca 72

<210> 231

<211> 72

<212> DNA

<213> Intelligent people

<400> 231

gacctcgtgg cgcaacggca gcgcgtctga ctccagatca gaaggttgcg tgttcaaatc 60

acgtcggggt ca 72

<210> 232

<211> 93

<212> DNA

<213> Intelligent people

<400> 232

ccttcaatag ttcagctggt agagcagagg actatagcta cttcctcagt aggagacgtc 60

cttaggttgc tggttcgatt ccagcttgaa gga 93

<210> 233

<211> 91

<212> DNA

<213> Intelligent people

<400> 233

ccttcgatag ctcagttggt agagcggagg actgtagttg gctgtgtcct tagacatcct 60

taggtcgctg gttcgaatcc ggctcgaagg a 91

<210> 234

<211> 89

<212> DNA

<213> Intelligent people

<400> 234

ccttcgatag ctcagttggt agagcggagg actgtagtgg atagggcgtg gcaatcctta 60

ggtcgctggt tcgattccgg ctcgaagga 89

<210> 235

<211> 89

<212> DNA

<213> Intelligent people

<400> 235

ccttcgatag ctcagttggt agagcggagg actgtaggct cattaagcaa ggtatcctta 60

ggtcgctggt tcgaatccgg ctcggagga 89

<210> 236

<211> 94

<212> DNA

<213> Intelligent people

<400> 236

ccttcgatag ctcagctggt agagcggagg actgtagatt gtatagacat ttgcggacat 60

ccttaggtcg ctggttcgat tccagctcga agga 94

<210> 237

<211> 93

<212> DNA

<213> Intelligent people

<400> 237

ccttcgatag ctcagctggt agagcggagg actgtagcta cttcctcagc aggagacatc 60

cttaggtcgc tggttcgatt ccggctcgaa gga 93

<210> 238

<211> 89

<212> DNA

<213> Intelligent people

<400> 238

ccttcgatag ctcagctggt agagcggagg actgtaggcg cgcgcccgtg gccatcctta 60

ggtcgctggt tcgattccgg ctcgaagga 89

<210> 239

<211> 94

<212> DNA

<213> Intelligent people

<400> 239

ccttcgatag ctcagctggt agagcggagg actgtagcct gtagaaacat ttgtggacat 60

ccttaggtcg ctggttcgat tccggctcga agga 94

<210> 240

<211> 94

<212> DNA

<213> Intelligent people

<400> 240

ccttcgatag ctcagctggt agagcggagg actgtagatt gtacagacat ttgcggacat 60

ccttaggtcg ctggttcgat tccggctcga agga 94

<210> 241

<211> 89

<212> DNA

<213> Intelligent people

<400> 241

ccttcgatag ctcagctggt agagcggagg actgtagtac ttaatgtgtg gtcatcctta 60

ggtcgctggt tcgattccgg ctcgaagga 89

<210> 242

<211> 89

<212> DNA

<213> Intelligent people

<400> 242

ccttcgatag ctcagctggt agagcggagg actgtagggg tttgaatgtg gtcatcctta 60

ggtcgctggt tcgaatccgg ctcggagga 89

<210> 243

<211> 94

<212> DNA

<213> Intelligent people

<400> 243

ccttcgatag ctcagctggt agagcggagg actgtagact gcggaaacgt ttgtggacat 60

ccttaggtcg ctggttcaat tccggctcga agga 94

<210> 244

<211> 90

<212> DNA

<213> Intelligent people

<400> 244

ctttcgatag ctcagttggt agagcggagg actgtaggtt cattaaacta aggcatcctt 60

aggtcgctgg ttcgaatccg gctcgaagga 90

<210> 245

<211> 88

<212> DNA

<213> Intelligent people

<400> 245

tcttcaatag ctcagctggt agagcggagg actgtaggtg cacgcccgtg gccattctta 60

ggtgctggtt tgattccgac ttggagag 88

<210> 246

<211> 73

<212> DNA

<213> Intelligent people

<400> 246

gtttccgtag tgtagtggtt atcacgttcg cctaacacgc gaaaggtccc cggttcgaaa 60

ccgggcggaa aca 73

<210> 247

<211> 73

<212> DNA

<213> Intelligent people

<400> 247

gtttccgtag tgtagtggtc atcacgttcg cctaacacgc gaaaggtccc cggttcgaaa 60

ccgggcggaa aca 73

<210> 248

<211> 73

<212> DNA

<213> Intelligent people

<400> 248

gtttccgtag tgtagtggtt atcacgttcg cctaacacgc gaaaggtccc tggatcaaaa 60

ccaggcggaa aca 73

<210> 249

<211> 73

<212> DNA

<213> Intelligent people

<400> 249

gtttccgtag tgtagtggtt atcacgttcg cctaacacgc gaaaggtccg cggttcgaaa 60

ccgggcggaa aca 73

<210> 250

<211> 73

<212> DNA

<213> Intelligent people

<400> 250

gtttccgtag tgtagtggtt atcacgtttg cctaacacgc gaaaggtccc cggttcgaaa 60

ccgggcagaa aca 73

<210> 251

<211> 72

<212> DNA

<213> Intelligent people

<400> 251

gggggtgtag ctcagtggta gagcgtatgc ttaacattca tgaggctctg ggttcgatcc 60

ccagcacttc ca 72

<210> 252

<211> 73

<212> DNA

<213> Intelligent people

<400> 252

gtttccgtag tgtagtggtt atcacgttcg cctcacacgc gaaaggtccc cggttcgaaa 60

ccgggcggaa aca 73

<210> 253

<211> 73

<212> DNA

<213> Intelligent people

<400> 253

gcttctgtag tgtagtggtt atcacgttcg cctcacacgc gaaaggtccc cggttcgaaa 60

ccgggcagaa gca 73

<210> 254

<211> 73

<212> DNA

<213> Intelligent people

<400> 254

gtttccgtag tgtagcggtt atcacattcg cctcacacgc gaaaggtccc cggttcgatc 60

ccgggcggaa aca 73

<210> 255

<211> 73

<212> DNA

<213> Intelligent people

<400> 255

gtttccgtag tgtagtggtt atcacgttcg cctcacacgc gaaaggtccc cggttcgaaa 60

ctgggcggaa aca 73

<210> 256

<211> 74

<212> DNA

<213> Intelligent people

<400> 256

gtttccgtag tgtagtggtt atcacgttcg cctcacacgc gtaaaggtcc ccggttcgaa 60

accgggcgga aaca 74

<210> 257

<211> 73

<212> DNA

<213> Intelligent people

<400> 257

gtttccgtag tggagtggtt atcacgttcg cctcacacgc gaaaggtccc cggtttgaaa 60

ccaggcggaa aca 73

<210> 258

<211> 73

<212> DNA

<213> Intelligent people

<400> 258

ggttccatag tgtagtggtt atcacgtctg ctttacacgc agaaggtcct gggttcgagc 60

cccagtggaa cca 73

<210> 259

<211> 73

<212> DNA

<213> Intelligent people

<400> 259

ggttccatag tgtagcggtt atcacgtctg ctttacacgc agaaggtcct gggttcgagc 60

cccagtggaa cca 73

<210> 260

<211> 73

<212> DNA

<213> Intelligent people

<400> 260

ggttccatag tgtagtggtt atcacatctg ctttacacgc agaaggtcct gggttcaagc 60

cccagtggaa cca 73

<210> 261

<211> 73

<212> DNA

<213> Intelligent people

<400> 261

gtttccgtgg tgtagtggtt atcacattcg ccttacacgc gaaaggtcct cgggtcgaaa 60

ccgagcggaa aca 73

<210> 262

<211> 72

<212> DNA

<213> Intelligent people

<400> 262

agcagagtgg cgcagcggaa gcgtgctggg cccataaccc agaggtcgat ggatcgaaac 60

catcctctgc ta 72

<210> 263

<211> 72

<212> DNA

<213> Intelligent people

<400> 263

agcagagtgg cgcagcggaa gcgtgctggg cccataaccc agaggtcgat ggatctaaac 60

catcctctgc ta 72

<210> 264

<211> 73

<212> DNA

<213> Intelligent people

<400> 264

tccctggtgg tctagtggct aggattcggc gctttcaccg ccgcggcccg ggttcgattc 60

ccggtcaggg aat 73

<210> 265

<211> 72

<212> DNA

<213> Intelligent people

<400> 265

gcgttggtgg tttagtggta gaattctcgc ctcccatgcg ggagacccgg gttcaattcc 60

cggccactgc ac 72

<210> 266

<211> 72

<212> DNA

<213> Intelligent people

<400> 266

ggccttggtg gtgcagtggt agaattctcg cctcccacgt gggagacccg ggttcaattc 60

ccggccaatg ca 72

<210> 267

<211> 73

<212> DNA

<213> Intelligent people

<400> 267

gtccctggtg gtctagtggc taggattcgg cgctttcacc gccgcggccc gggttcgatt 60

cccggccagg gaa 73

<210> 268

<211> 75

<212> DNA

<213> Intelligent people

<400> 268

tgtctctgtg gcgcaatcgg ttagcgcgtt cggctgttaa ccgaaagatt ggtggttcga 60

gcccacccag ggacg 75

<210> 269

<211> 86

<212> DNA

<213> Intelligent people

<400> 269

tggctccgtg gcgcaatgga tagcgcattg gacttctaga ggctgaaggc attcaaaggt 60

tccgggttcg agtcccggcg gagtcg 86

<210> 270

<211> 74

<212> DNA

<213> Intelligent people

<400> 270

gcccggctag ctcagtcggt agagcatgag actcttaatc tcagggtcgt gggttcgagc 60

cccacgttgg gcgc 74

<210> 271

<211> 73

<212> DNA

<213> Intelligent people

<400> 271

gccgtgatcg tatagtggtt agtactctgc gttgtggccg cagcaacctc ggttcgaatc 60

cgagtcacgg cag 73

<210> 272

<211> 73

<212> DNA

<213> Intelligent people

<400> 272

gcgttggtgg tatagtggtg agcatagctg ccttccaagc agttgacccg ggttcgattc 60

ccggccaacg cag 73

<210> 273

<211> 73

<212> DNA

<213> Intelligent people

<400> 273

tccctggtgg tctagtggtt aggattcggc gctctcaccg ccgcggcccg ggttcgattc 60

ccggtcaggg aaa 73

<210> 274

<211> 73

<212> DNA

<213> Intelligent people

<400> 274

aggttccatg gtgtaatggt gagcactctg gactctgaat ccagcgatcc gagttcgagt 60

ctcggtggaa cct 73

<210> 275

<211> 75

<212> DNA

<213> Intelligent people

<400> 275

tgtctctgtg gcgtagtcgg ttagcgcgtt cggctgttaa ccgaaaagtt ggtggttcga 60

gcccacccag gaacg 75

<210> 276

<211> 75

<212> DNA

<213> Intelligent people

<400> 276

tgtctctgtg gcgcaatcgg ttagcgcgtt cggctgttaa ccgaaaggtt ggtggttcga 60

gcccacccag ggacg 75

<210> 277

<211> 75

<212> DNA

<213> Intelligent people

<400> 277

gtctctgtgg cgcaatcggt tagcgcattc ggctgttaac cgaaaggttg gtggttcgag 60

cccacccagg gacgc 75

<210> 278

<211> 75

<212> DNA

<213> Intelligent people

<400> 278

gtctctgtgg cgcaatgggt tagcgcgttc ggctgttaac cgaaaggttg gtggttcgag 60

cccatccagg gacgc 75

<210> 279

<211> 72

<212> DNA

<213> Intelligent people

<400> 279

gcactggtgg ttcagtggta gaattctcgc ctcacacgcg ggacacccgg gttcaattcc 60

cggtcaaggc aa 72

<210> 280

<211> 74

<212> DNA

<213> Intelligent people

<400> 280

gtttccgtag tgtagtggtt atcacgttcg cctcacacgc gaaaggtccc cggttcgaaa 60

ctgggcggaa acag 74

<210> 281

<211> 72

<212> DNA

<213> Intelligent people

<400> 281

gcactggtgg ttcagtggta gaattctcgc ctcccacgcg ggagacccgg gtttaattcc 60

cggtcaagat aa 72

<210> 282

<211> 75

<212> DNA

<213> Intelligent people

<400> 282

gtttccgtag tgtagtggtt atcacgttcg cctcacacgc gtaaaggtcc ccggttcgaa 60

accgggcgga aacat 75

<210> 283

<211> 73

<212> DNA

<213> Intelligent people

<400> 283

tagcagagtg gcgcagcgga agcgtgctgg gcccataacc cagaggtcga tggatcgaaa 60

ccatcctctg cta 73

<210> 284

<211> 74

<212> DNA

<213> Intelligent people

<400> 284

gtttccgtag tgtagtggtt atcacgttcg cctcacacgc gaaaggtccc cggttcgaaa 60

ccgggcggaa acaa 74

<210> 285

<211> 73

<212> DNA

<213> Intelligent people

<400> 285

tcctcgttag tatagtggtg agtatccccg cctgtcacgc gggagaccgg ggttcgattc 60

cccgacgggg agg 73

<210> 286

<211> 72

<212> DNA

<213> Intelligent people

<400> 286

tgcatgggtg gttcagtggt agaattctcg cctgccacgc gggaggcccg ggttcgattc 60

ccggcccatg ca 72

<210> 287

<211> 73

<212> DNA

<213> Intelligent people

<400> 287

tccctggtgg tctagtggtt aggattcggc gctctcaccg ccgcggcccg ggttcgattc 60

ccggtcaggg aag 73

<210> 288

<211> 73

<212> DNA

<213> Intelligent people

<400> 288

atccttgtta ctatagtggt gagtatctct gcctgtcatg cgtgagagag ggggtcgatt 60

ccccgacggg gag 73

<210> 289

<211> 72

<212> DNA

<213> Intelligent people

<400> 289

gcattggtgg ttcagtggta gaattctcgc ctgccacgcg ggaggcccgg gttcgattcc 60

cggccaatgc ac 72

<210> 290

<211> 84

<212> DNA

<213> Intelligent people

<400> 290

gtcaggatgg ccgagcggtc taaggcgctg cgttcaggtc gcagtctccc ctggaggcgt 60

gggttcgaat cccactcctg acaa 84

<210> 291

<211> 73

<212> DNA

<213> Intelligent people

<400> 291

cgcgttggtg gtatagtggt gagcatagct gccttccaag cagttgaccc gggttcgatt 60

cccggccaac gca 73

<210> 292

<211> 75

<212> DNA

<213> Intelligent people

<400> 292

cgtctctgtg gcgcaatcgg ttagcgcgtt cggctgttaa ccgaaaggtt ggtggttcga 60

tcccacccag ggacg 75

<210> 293

<211> 73

<212> DNA

<213> Intelligent people

<400> 293

cgcgttggtg gtgtagtggt gagcacagct gcctttcaag cagttaacgc gggttcgatt 60

cccgggtaac gaa 73

<210> 294

<211> 73

<212> DNA

<213> Intelligent people

<400> 294

cggctcgttg gtctaggggt atgattctcg cttcgggtgc gagaggtccc gggttcaaat 60

cccggacgag ccc 73

<210> 295

<211> 73

<212> DNA

<213> Intelligent people

<400> 295

ggctcgttgg tctaggggta tgattctcgc ttagggtgcg agaggtcccg ggttcaaatc 60

ccggacgagc cct 73

<210> 296

<211> 74

<212> DNA

<213> Intelligent people

<400> 296

cgcccggata gctcagtcgg tagagcatca gacttttaat ctgagggtcc agggttcaag 60

tccctgttcg ggcg 74

<210> 297

<211> 74

<212> DNA

<213> Intelligent people

<400> 297

gcccggatag ctcagtcggt agagcatcag acttttaatc tgagggtcca gggttcaagt 60

ccctgttcgg gcgt 74

<210> 298

<211> 107

<212> DNA

<213> Intelligent people

<400> 298

tgtcaggatg gccgagtggt ctaaggcgcc agactcaagg taagcacctt gcctgcgggc 60

tttctggtct ccggatggag gcgtgggttc gaatcccact tctgaca 107

<210> 299

<211> 73

<212> DNA

<213> Intelligent people

<400> 299

ttccctggtg gtctagtggt taggattcgg cgctctcacc gccgcggccc gggttcgatt 60

cccggtcagg aaa 73

<210> 300

<211> 90

<212> DNA

<213> Intelligent people

<400> 300

gccttcgata gctcagttgg tagagcggag gactgtagtg gatagggcgt ggcaatcctt 60

aggtcgctgg ttcgattccg gctcgaagga 90

<210> 301

<211> 74

<212> DNA

<213> Intelligent people

<400> 301

cgggggatta gctcaaatgg tagagcgctc gcttagcatg cgagaggtag cgggatcgat 60

gcccgcatcc tcca 74

<210> 302

<211> 94

<212> DNA

<213> Intelligent people

<400> 302

agctccagtg gcgcaatcgg ttagcgcgcg gtacttatac agcagtacat gcagagcaat 60

gccgaggttg tgagttcgag cctcacctgg agca 94

<210> 303

<211> 72

<212> DNA

<213> Intelligent people

<400> 303

gcgccgctgg tgtagtggta tcatgcaaga ttcccattct tgcgacccgg gttcgattcc 60

cgggcggcgc at 72

<210> 304

<211> 73

<212> DNA

<213> Intelligent people

<400> 304

tcccatatgg tctagcggtt aggattcctg gttttcaccc aggtggcccg ggttcgactc 60

ccggtatggg aac 73

<210> 305

<211> 73

<212> DNA

<213> Intelligent people

<400> 305

gggggatgta gctcagtggt agagcgcgcg cttcgcatgt gtgaggtccc gggttcaatc 60

cccggcatct cca 73

<210> 306

<211> 72

<212> DNA

<213> Intelligent people

<400> 306

gcattggtgg ttcagtggta gaattctcgc ctgccacgcg ggaggcccgg gttcgattcc 60

cggccaatgc aa 72

<210> 307

<211> 74

<212> DNA

<213> Intelligent people

<400> 307

gggccagtgg cgcaatggat aacgcgtctg actacggatc agaagattct aggttcgact 60

cctggctggc tcgc 74

<210> 308

<211> 74

<212> DNA

<213> Intelligent people

<400> 308

ggtttccgta gtgtagtggt tatcacgttc gcctaacacg cgaaaggtcc ccggttcgaa 60

accgggcgga aaca 74

<210> 309

<211> 74

<212> DNA

<213> Intelligent people

<400> 309

agtttccgta gtgtagtggt tatcacgttc gcctaacacg cgaaaggtcc ccggttcgaa 60

accgggcgga aaca 74

<210> 310

<211> 83

<212> DNA

<213> Intelligent people

<400> 310

aggtagcgtg gccgagcggt ctaaggcgct ggattaaggc tccagtctct tcgggggcgt 60

gggttcgaat cccaccgctg cca 83

<210> 311

<211> 74

<212> DNA

<213> Intelligent people

<400> 311

gtttccgtag tgtagtggtc atcacgttcg cctaacacgc gaaaggtccc cggttcgaaa 60

ccgggcggaa acat 74

<210> 312

<211> 73

<212> DNA

<213> Intelligent people

<400> 312

ggctcgttgg tctaggggta tgattctcgc tttgggtgcg agaggtcccg ggttcaaatc 60

ccggacgagc cca 73

<210> 313

<211> 73

<212> DNA

<213> Intelligent people

<400> 313

ggctccatag ctcaggggtt agagcactgg tcttgtaaac cagggtcgcg agttcaaatc 60

tcgctggggc ctg 73

<210> 314

<211> 73

<212> DNA

<213> Intelligent people

<400> 314

tggggatgta gctcagtggt agagcgcatg ctttgcatgt atgaggcccc gggttcgatc 60

cccggcatct cca 73

<210> 315

<211> 74

<212> DNA

<213> Intelligent people

<400> 315

cgcccggcta gctcagtcgg tagagcatga gactcttaat ctcagggtcg tgggttcgag 60

ccccacgttg ggcg 74

<210> 316

<211> 74

<212> DNA

<213> Intelligent people

<400> 316

gtttccgtag tgtagtggtt atcacgttcg cctaacacgc gaaaggtccc cggttcgaaa 60

ccgggcggaa acaa 74

<210> 317

<211> 74

<212> DNA

<213> Intelligent people

<400> 317

gcccggctag ctcagtcggt agagcatgag actcttaatc tcagggtcgt gggttcgagc 60

cccacgttgg gcgt 74

<210> 318

<211> 74

<212> DNA

<213> Intelligent people

<400> 318

gtttccgtag tgtagtggtt atcacgttcg cctcacacgc gaaaggtccc cggttcgaaa 60

ccgggcggaa acac 74

<210> 319

<211> 73

<212> DNA

<213> Intelligent people

<400> 319

cagcagagtg gcgcagcgga agcgtgctgg gcccataacc cagaggtcga tggatcgaaa 60

ccatcctctg cta 73

<210> 320

<211> 85

<212> DNA

<213> Intelligent people

<400> 320

ggagaggcct ggccgagtgg ttaaggcgat ggactgctaa tccattgtgc tctgcacgcg 60

tgggttcgaa tcccatcctc gtcgc 85

<210> 321

<211> 73

<212> DNA

<213> Intelligent people

<400> 321

ggccccatgg tgtaatggtt agcactctgg actttgaatc cagcgatccg agttcaaatc 60

tcggtgggac ctg 73

<210> 322

<211> 73

<212> DNA

<213> Intelligent people

<400> 322

ggccccatgg tgtaatggtt agcactctgg actttgaatc cagcgatccg agttcaaatc 60

tcggtgggac cta 73

<210> 323

<211> 83

<212> DNA

<213> Intelligent people

<400> 323

gtagtcgtgg ccgagtggtt aaggcgatgg acttgaaatc cattggggtc tccccgcgca 60

ggttcgaatc ctgccgacta cgg 83

<210> 324

<211> 73

<212> DNA

<213> Intelligent people

<400> 324

agcagagtgg cgcagcggaa gcgtgctggg cccataaccc agaggtcgat ggatcgaaac 60

catcctctgc tat 73

<210> 325

<211> 74

<212> DNA

<213> Intelligent people

<400> 325

ggaccacgtg gcctaatgga taaggcgtct gacttcggat cagaagattg agggttcgaa 60

tccctccgtg gtta 74

<210> 326

<211> 83

<212> DNA

<213> Intelligent people

<400> 326

tgtagtcgtg gccgagtggt taaggcgatg gactagaaat ccattggggt ctccccgcgc 60

aggttcgaat cctgccgact acg 83

<210> 327

<211> 73

<212> DNA

<213> Intelligent people

<400> 327

agcagagtgg cgcagcggaa gcgtgctggg cccataaccc agaggtcgat ggatcgaaac 60

catcctctgc tag 73

<210> 328

<211> 84

<212> DNA

<213> Intelligent people

<400> 328

cgtcaggatg gccgagcggt ctaaggcgct gcgttcaggt cgcagtctcc cctggaggcg 60

tgggttcgaa tcccactcct gaca 84

<210> 329

<211> 75

<212> DNA

<213> Intelligent people

<400> 329

ggctccgtgg cttagctggt taaagcgcct gtctagtaaa caggagatcc tgggttcgaa 60

tcccagcggg gcctg 75

<210> 330

<211> 74

<212> DNA

<213> Intelligent people

<400> 330

agggccagtg gcgcaatgga taacgcgtct gactacggat cagaagattc caggttcgac 60

tcctggctgg ctcg 74

<210> 331

<211> 74

<212> DNA

<213> Intelligent people

<400> 331

ggtttccgta gtgtagtggt tatcacgttc gcctcacacg cgaaaggtcc ccggttcgaa 60

accgggcgga aaca 74

<210> 332

<211> 73

<212> DNA

<213> Intelligent people

<400> 332

aggggatgta gctcagtggt agagcgcatg cttcgcatgt atgaggtccc gggttcgatc 60

cccggcatct cca 73

<210> 333

<211> 75

<212> DNA

<213> Intelligent people

<400> 333

tggccggtta gctcagttgg ttagagcgtg gtgctaataa cgccaaggtc gcgggttcga 60

tccccgtacg ggcca 75

<210> 334

<211> 73

<212> DNA

<213> Intelligent people

<400> 334

cggctcgttg gtctaggggt atgattctcg cttagggtgc gagaggtccc gggttcaaat 60

cccggacgag ccc 73

<210> 335

<211> 74

<212> DNA

<213> Intelligent people

<400> 335

agcccggcta gctcagtcgg tagagcatga gactcttaat ctcagggtcg tgggttcgag 60

ccccacgttg ggcg 74

<210> 336

<211> 92

<212> DNA

<213> Intelligent people

<400> 336

tccttcgata gctcagttgg tagagcggag gactgtagtt ggctgtgtcc ttagacatcc 60

ttaggtcgct ggttcgaatc cggctcgaag ga 92

<210> 337

<211> 74

<212> DNA

<213> Intelligent people

<400> 337

ggggaattag ctcaaatggt agagcgctcg cttagcatgc gagaggtagc gggatcgatg 60

cccgcattct ccag 74

<210> 338

<211> 74

<212> DNA

<213> Intelligent people

<400> 338

cgccctctta gcgcagcggg cagcgcgtca gtctcataat ctgaaggtcc tgagttcgag 60

cctcagagag ggca 74

<210> 339

<211> 95

<212> DNA

<213> Intelligent people

<400> 339

tgctccagtg gcgcaatcgg ttagcgcgcg gtacttatat ggcagtatgt gtgcgagtga 60

tgccgaggtt gtgagttcga gcctcacctg gagca 95

<210> 340

<211> 73

<212> DNA

<213> Intelligent people

<400> 340

tgccgtgatc gtatagtggt tagtactctg cgttgtggcc gcagcaacct cggttcgaat 60

ccgagtcacg gca 73

<210> 341

<211> 75

<212> DNA

<213> Intelligent people

<400> 341

ggccggttag ctcagttggt tagagcgtgg tgctaataac gccaaggtcg cgggttcgat 60

ccccgtacgg gccac 75

<210> 342

<211> 74

<212> DNA

<213> Intelligent people

<400> 342

agtttccgta gtgtagtggt tatcacgttt gcctaacacg cgaaaggtcc ccggttcgaa 60

accgggcaga aaca 74

<210> 343

<211> 74

<212> DNA

<213> Intelligent people

<400> 343

gcttctgtag tgtagtggtt atcacgttcg cctcacacgc gaaaggtccc cggttcgaaa 60

ccgggcagaa gcaa 74

<210> 344

<211> 73

<212> DNA

<213> Intelligent people

<400> 344

ttcctcgtta gtatagtggt gagtatcccc gcctgtcacg cgggagaccg gggttcgatt 60

ccccgacggg gag 73

<210> 345

<211> 83

<212> DNA

<213> Intelligent people

<400> 345

gtagtcgtgg ccgagtggtt aaggcgatgg acttgaaatc cattggggtt tccccgcgca 60

ggttcgaatc ctgtcggcta cgg 83

<210> 346

<211> 73

<212> DNA

<213> Intelligent people

<400> 346

aggttccatg gtgtaatggt tagcactctg gactctgaat ccagcgatcc gagttcaaat 60

ctcggtggaa cct 73

<210> 347

<211> 73

<212> DNA

<213> Intelligent people

<400> 347

tcctcgttag tatagtggtg agtgtccccg tctgtcacgc gggagaccgg ggttcgattc 60

cccgacgggg aga 73

<210> 348

<211> 74

<212> DNA

<213> Intelligent people

<400> 348

gtttccgtag tgtagtggtt atcacgttcg cctaacacgc gaaaggtccc tggatcaaaa 60

ccaggcggaa acaa 74

<210> 349

<211> 75

<212> DNA

<213> Intelligent people

<400> 349

cggccggtta gctcagttgg ttagagcgtg gtgctaataa cgccaaggtc gcgggttcga 60

tccccgtact ggcca 75

<210> 350

<211> 73

<212> DNA

<213> Intelligent people

<400> 350

ggccccatgg tgtaatggtc agcactctgg actctgaatc cagcgatccg agttcaaatc 60

tcggtgggac cca 73

<210> 351

<211> 73

<212> DNA

<213> Intelligent people

<400> 351

ggccccatgg tgtaatggtt agcactctgg actttgaatc cagcgatccg agttcaaatc 60

tcggtgggac ctt 73

<210> 352

<211> 73

<212> DNA

<213> Intelligent people

<400> 352

tgggggtgta gctcagtggt agagcgcgtg cttagcatgt acgaggtccc gggttcaatc 60

cccggcacct cca 73

<210> 353

<211> 73

<212> DNA

<213> Intelligent people

<400> 353

ggggatgtag ctcagtggta gagcgcatgc ttagcatgca tgaggtcccg ggttcgatcc 60

ccagcatctc cag 73

<210> 354

<211> 73

<212> DNA

<213> Intelligent people

<400> 354

agggggtgta gctcagtggt agagcgcgtg cttcgcatgt acgaggcccc gggttcgacc 60

cccggctcct cca 73

<210> 355

<211> 73

<212> DNA

<213> Intelligent people

<400> 355

gggggtgtag ctcagtggta gagcgcgtgc ttagcatgca cgaggccccg ggttcaatcc 60

ccggcacctc cat 73

<210> 356

<211> 73

<212> DNA

<213> Intelligent people

<400> 356

gggggtgtag ctcagtggta gagcgcgtgc ttagcatgca cgaggccccg ggttcaatcc 60

ccggcacctc cag 73

<210> 357

<211> 107

<212> DNA

<213> Intelligent people

<400> 357

gtcaggatgg ccgagtggtc taaggcgcca gactcaagct aagcttcctc cgcggtgggg 60

attctggtct ccaatggagg cgtgggttcg aatcccactt ctgacac 107

<210> 358

<211> 106

<212> DNA

<213> Intelligent people

<400> 358

tgtcaggatg gccgagtggt ctaaggcgcc agactcaagc ttggcttcct cgtgttgagg 60

attctggtct ccaatggagg cgtgggttcg aatcccactt ctgaca 106

<210> 359

<211> 73

<212> DNA

<213> Intelligent people

<400> 359

ggttccatgg tgtaatggtt agcactctgg actctgaatc cagcgatccg agttcaaatc 60

tcggtggaac ctt 73

<210> 360

<211> 83

<212> DNA

<213> Intelligent people

<400> 360

ggtagcgtgg ccgagcggtc taaggcgctg gattaaggct ccagtctctt cgggggcgtg 60

ggttcgaatc ccaccgctgc cag 83

<210> 361

<211> 74

<212> DNA

<213> Intelligent people

<400> 361

tgcctcctta gcgcagtagg cagcgcgtca gtctcataat ctgaaggtcc tgagttcgaa 60

cctcagaggg ggca 74

<210> 362

<211> 74

<212> DNA

<213> Intelligent people

<400> 362

agcccggata gctcagtcgg tagagcatca gacttttaat ctgagggtcc agggttcaag 60

tccctgttcg ggcg 74

<210> 363

<211> 74

<212> DNA

<213> Intelligent people

<400> 363

gcctccttag cgcagtaggc agcgcgtcag tctcataatc tgaaggtcct gagttcgaac 60

ctcagagggg gcag 74

<210> 364

<211> 73

<212> DNA

<213> Intelligent people

<400> 364

ttccctggtg gtctagtggt taggattcgg cgctctcacc gccgcggccc gggttcgatt 60

cccggtcagg gaa 73

<210> 365

<211> 84

<212> DNA

<213> Intelligent people

<400> 365

caccaggatg gccgagtggt taaggcgttg gacttaagat ccaatggaca tatgtccgcg 60

tgggttcgaa ccccactcct ggta 84

<210> 366

<211> 73

<212> DNA

<213> Intelligent people

<400> 366

tggctcgttg gtctaggggt atgattctcg cttagggtgc gagaggtccc gggttcaaat 60

cccggacgag ccc 73

<210> 367

<211> 74

<212> DNA

<213> Intelligent people

<400> 367

agccccagtg gcctaatgga taaggcattg gcctcctaag ccagggattg tgggttcgag 60

tcccatctgg ggtg 74

<210> 368

<211> 73

<212> DNA

<213> Intelligent people

<400> 368

ggggatatag ctcaggggta gagcatttga ctgcagatca agaggtcccc ggttcaaatc 60

cgggtgcccc ccc 73

<210> 369

<211> 94

<212> DNA

<213> Intelligent people

<400> 369

cccttcgata gctcagctgg tagagcggag gactgtagct acttcctcag caggagacat 60

ccttaggtcg ctggttcgat tccggctcga agga 94

<210> 370

<211> 90

<212> DNA

<213> Intelligent people

<400> 370

cccttcgata gctcagctgg tagagcggag gactgtaggc gcgcgcccgt ggccatcctt 60

aggtcgctgg ttcgattccg gctcgaagga 90

<210> 371

<211> 74

<212> DNA

<213> Intelligent people

<400> 371

tgggggatta gctcaaatgg tagagcgctc gcttagcatg cgagaggtag cgggatcgat 60

gcccgcatcc tcca 74

<210> 372

<211> 83

<212> DNA

<213> Intelligent people

<400> 372

gtagtcgtgg ccgagtggtt aaggcgatgg actagaaatc cattggggtc tccccgcgca 60

ggttcgaatc ctgccgacta cgg 83

<210> 373

<211> 74

<212> DNA

<213> Intelligent people

<400> 373

gcctcgttag cgcagtaggt agcgcgtcag tctcataatc tgaaggtcgt gagttcgatc 60

ctcacacggg gcac 74

<210> 374

<211> 92

<212> DNA

<213> Intelligent people

<400> 374

ggctctgtgg cgcaatggat agcgcattgg acttctagct gagcctagtg tggtcattca 60

aaggttgtgg gttcgagtcc caccagagtc ga 92

<210> 375

<211> 75

<212> DNA

<213> Intelligent people

<400> 375

gtctctgtgg cgcaatcggt tagcgcgttc ggctgttaac cgaaaggttg gtggttcgag 60

cccacccagg gacgc 75

<210> 376

<211> 83

<212> DNA

<213> Intelligent people

<400> 376

ggcagcgatg gccgagtggt taaggcgttg gacttgaaat ccaatggggt ctccccgcgc 60

aggttcgaac cctgctcgct gcg 83

<210> 377

<211> 74

<212> DNA

<213> Intelligent people

<400> 377

ggttccatag tgtagtggtt atcacgtctg ctttacacgc agaaggtcct gggttcgagc 60

cccagtggaa ccat 74

<210> 378

<211> 74

<212> DNA

<213> Intelligent people

<400> 378

ggttccatag tgtagcggtt atcacgtctg ctttacacgc agaaggtcct gggttcgagc 60

cccagtggaa ccac 74

<210> 379

<211> 87

<212> DNA

<213> Intelligent people

<400> 379

tggctctgtg gcgcaatgga tagcgcattg gacttctaga tagttagaga aattcaaagg 60

ttgtgggttc gagtcccacc agagtcg 87

<210> 380

<211> 84

<212> DNA

<213> Intelligent people

<400> 380

taccagaatg gccgagtggt taaggcgttg gacttaagat ccaatggatt catatccgcg 60

tgggttcgaa ccccacttct ggta 84

<210> 381

<211> 74

<212> DNA

<213> Intelligent people

<400> 381

ggcccggata gctcagtcgg tagagcatca gacttttaat ctgagggtcc ggggttcaag 60

tccctgttcg ggcg 74

<210> 382

<211> 74

<212> DNA

<213> Intelligent people

<400> 382

gccgaaatag ctcagttggg agagcgttag actgaagatc taaaggtccc tggttcgatc 60

ccgggtttcg gcag 74

<210> 383

<211> 74

<212> DNA

<213> Intelligent people

<400> 383

gcccggatag ctcagtcggt agagcatcag acttttaatc tgagggtcca gggttcaagt 60

ccctgttcgg gcgg 74

<210> 384

<211> 74

<212> DNA

<213> Intelligent people

<400> 384

gccgaaatag ctcagttggg agagcgttag actgaagatc taaaggtccc tggttcaatc 60

ccgggtttcg gcag 74

<210> 385

<211> 83

<212> DNA

<213> Intelligent people

<400> 385

ggacgaggtg gccgagtggt taaggcgatg gactgctaat ccattgtgct ttgcacgcgt 60

gggttcgaat cccatcctcg tcg 83

<210> 386

<211> 73

<212> DNA

<213> Intelligent people

<400> 386

ggctcgttgg tctaggggta tgattctcgg tttgggtccg agaggtcccg ggttcaaatc 60

ccggacgagc ccc 73

<210> 387

<211> 83

<212> DNA

<213> Intelligent people

<400> 387

agtcacggtg gccgagtggt taaggcgttg gactcgaaat ccaatggggt ttccccgcac 60

aggttcgaat cctgttcgtg acg 83

<210> 388

<211> 73

<212> DNA

<213> Intelligent people

<400> 388

ctcctcgtta gtatagtggt tagtatcccc gcctgtcacg cgggagaccg gggttcaatt 60

ccccgacggg gag 73

<210> 389

<211> 73

<212> DNA

<213> Intelligent people

<400> 389

ggacctcgtg gcgcaacggt agcgcgtctg actccagatc agaaggctgc gtgttcgaat 60

cacgtcgggg tca 73

<210> 390

<211> 73

<212> DNA

<213> Intelligent people

<400> 390

ggggatgtag ctcagtggta gagcgcatgc tttgcatgta tgaggccccg ggttcgatcc 60

ccggcatctc cat 73

<210> 391

<211> 74

<212> DNA

<213> Intelligent people

<400> 391

gccgaaatag ctcagttggg agagcgttag actgaagatc taaaggtccc tggttcgatc 60

ccgggtttcg gcac 74

<210> 392

<211> 73

<212> DNA

<213> Intelligent people

<400> 392

aggggatgta gctcagtggt agagcgcatg ctttgcacgt atgaggcccc gggttcaatc 60

cccggcatct cca 73

<210> 393

<211> 75

<212> DNA

<213> Intelligent people

<400> 393

gtctctgtgg cgcaatcggt tagcgcgttc ggctgttaac cgaaaggttg gtggttcgag 60

cccacccagg gacgg 75

<210> 394

<211> 73

<212> DNA

<213> Intelligent people

<400> 394

tcccacatgg tctagcggtt aggattcctg gttttcaccc aggcggcccg ggttcgactc 60

ccggtgtggg aac 73

<210> 395

<211> 74

<212> DNA

<213> Intelligent people

<400> 395

ggctccatag ctcaggggtt agagcgctgg tcttgtaaac caggggtcgc gagttcaatt 60

ctcgctgggg cctg 74

<210> 396

<211> 83

<212> DNA

<213> Intelligent people

<400> 396

tggtagtgtg gccgagcggt ctaaggcgct ggatttaggc tccagtctct tcgggggcgt 60

gggttcgaat cccaccactg cca 83

<210> 397

<211> 74

<212> DNA

<213> Intelligent people

<400> 397

ggctccatag ctcaggggtt agagcactgg tcttgtaaac caggggtcgc gagttcaaat 60

ctcgctgggg cctc 74

<210> 398

<211> 73

<212> DNA

<213> Intelligent people

<400> 398

tggctcgttg gtctagtggt atgattctcg ctttgggtgc gagaggtccc gggttcaaat 60

cccggacgag ccc 73

<210> 399

<211> 95

<212> DNA

<213> Intelligent people

<400> 399

ccttcgatag ctcagctggt agagcggagg actgtagatt gtacagacat ttgcggacat 60

ccttaggtcg ctggttcgat tccggctcga aggaa 95

<210> 400

<211> 74

<212> DNA

<213> Intelligent people

<400> 400

aggccctata gctcaggggt tagagcactg gtcttgtaaa ccaggggtcg cgagttcaaa 60

tctcgctggg gcct 74

<210> 401

<211> 90

<212> DNA

<213> Intelligent people

<400> 401

tccttcgata gctcagctgg tagagcggag gactgtagta cttaatgtgt ggtcatcctt 60

aggtcgctgg ttcgattccg gctcgaagga 90

<210> 402

<211> 73

<212> DNA

<213> Intelligent people

<400> 402

tggctcgttg gtctaggggt atgattctcg ctttgggtgc gagaggtccc gggttcaaat 60

cccggacgag ccc 73

<210> 403

<211> 74

<212> DNA

<213> Intelligent people

<400> 403

gcccggctag ctcagtcggt agagcatggg actcttaatc ccagggtcgt gggttcgagc 60

cccacgttgg gcgc 74

<210> 404

<211> 75

<212> DNA

<213> Intelligent people

<400> 404

cggccggtta gctcagttgg ttagagcgtg gtgctaataa cgccaaggtc gcgggttcga 60

tccccgtacg ggcca 75

<210> 405

<211> 73

<212> DNA

<213> Intelligent people

<400> 405

tcccacatgg tctagcggtt aggattcctg gttttcaccc aggcggcccg ggttcgactc 60

ccggtgtggg aat 73

<210> 406

<211> 83

<212> DNA

<213> Intelligent people

<400> 406

gacgaggtgg ccgagtggtt aaggcgatgg actgctaatc cattgtgctc tgcacgcgtg 60

ggttcgaatc ccatcctcgt cga 83

<210> 407

<211> 73

<212> DNA

<213> Intelligent people

<400> 407

gccgtgatcg tatagtggtt agtactctgc gttgtggccg cagcaacctc ggttcgaatc 60

cgagtcacgg cat 73

<210> 408

<211> 73

<212> DNA

<213> Intelligent people

<400> 408

cgccgtgatc gtatagtggt tagtactctg cgttgtggcc gcagcaacct cggttcgaat 60

ccgagtcacg gca 73

<210> 409

<211> 73

<212> DNA

<213> Intelligent people

<400> 409

ggttccatgg tgtaatggtt agcactctgg actctgaatc cagcgatccg agttcaaatc 60

tcggtggaac ctg 73

<210> 410

<211> 74

<212> DNA

<213> Intelligent people

<400> 410

tgcccggcta gctcagtcgg tagagcatgg gactcttaat cccagggtcg tgggttcgag 60

ccccacgttg ggcg 74

<210> 411

<211> 74

<212> DNA

<213> Intelligent people

<400> 411

gggccgcgtg gcctaatgga taaggcgtct gacttcggat cagaagattg caggttcgag 60

tcctgccgcg gtcg 74

<210> 412

<211> 72

<212> DNA

<213> Intelligent people

<400> 412

gcgccgctgg tgtagtggta tcatgcaaga ttcccattct tgcgacccgg gttcgattcc 60

cgggcggcgc ac 72

<210> 413

<211> 74

<212> DNA

<213> Intelligent people

<400> 413

gggccgcgtg gcctaatgga taaggcgtct gattccggat cagaagattg agggttcgag 60

tcccttcgtg gtcg 74

<210> 414

<211> 74

<212> DNA

<213> Intelligent people

<400> 414

cgccccggtg gcctaatgga taaggcattg gcctcctaag ccagggattg tgggttcgag 60

tcccacccgg ggta 74

<210> 415

<211> 74

<212> DNA

<213> Intelligent people

<400> 415

gcccggctag ctcagtcggt agagcatgag acccttaatc tcagggtcgt gggttcgagc 60

cccacgttgg gcgt 74

<210> 416

<211> 73

<212> DNA

<213> Intelligent people

<400> 416

aggcgcggtg gccaagtggt aaggcgtcgg tctcgtaaac cgaagatcac gggttcgaac 60

cccgtccgtg cct 73

<210> 417

<211> 83

<212> DNA

<213> Intelligent people

<400> 417

ggtagcgtgg ccgagtggtc taaggcgctg gatttaggct ccagtcattt cgatggcgtg 60

ggttcgaatc ccaccgctgc cac 83

<210> 418

<211> 83

<212> DNA

<213> Intelligent people

<400> 418

gggtagcgtg gccgagcggt ctaaggcgct ggattaaggc tccagtctct tcgggggcgt 60

gggttcgaat cccaccgctg cca 83

<210> 419

<211> 84

<212> DNA

<213> Intelligent people

<400> 419

agtcaggatg gccgagcggt ctaaggcgct gcgttcaggt cgcagtctcc cctggaggcg 60

tgggttcgaa tcccacttct gaca 84

<210> 420

<211> 84

<212> DNA

<213> Intelligent people

<400> 420

gtcaggatgg ccgagcggtc taaggcgctg cgttcaggtc gcagtctccc ctggaggcgt 60

gggttcgaat cccacttctg acag 84

<210> 421

<211> 74

<212> DNA

<213> Intelligent people

<400> 421

gcctcgttag cgcagtaggc agcgcgtcag tctcataatc tgaaggtcgt gagttcgagc 60

ctcacacggg gcag 74

<210> 422

<211> 83

<212> DNA

<213> Intelligent people

<400> 422

ggtagcgtgg ccgagcggtc taaggcgctg gatttaggct ccagtctctt cggaggcgtg 60

ggttcgaatc ccaccgctgc cag 83

<210> 423

<211> 89

<212> DNA

<213> Intelligent people

<400> 423

tggctctgtg gcgcaatgga tagcgcattg gacttctagt gacgaataga gcaattcaaa 60

ggttgtgggt tcgaatccca ccagagtcg 89

<210> 424

<211> 72

<212> DNA

<213> Intelligent people

<400> 424

cgcattggtg gttcagtggt agaattctcg cctgccacgc gggaggcccg ggttcgattc 60

ccggccaatg ca 72

<210> 425

<211> 83

<212> DNA

<213> Intelligent people

<400> 425

gctgtgatgg ccgagtggtt aaggcgttgg actcgaaatc caatggggtc tccccgcgca 60

ggttcgaatc ctgctcacag cgt 83

<210> 426

<211> 75

<212> DNA

<213> Intelligent people

<400> 426

ggcgccgtgg cttagctggt taaagcgcct gtctagtaaa caggagatcc tgggttcgaa 60

tcccagcggt gcctg 75

<210> 427

<211> 73

<212> DNA

<213> Intelligent people

<400> 427

cgacctcgtg gcgcaacggt agcgcgtctg actccagatc agaaggttgc gtgttcaaat 60

cacgtcgggg tca 73

<210> 428

<211> 83

<212> DNA

<213> Intelligent people

<400> 428

agacgaggtg gccgagtggt taaggcgatg gactgctaat ccattgtgct ctgcacgcgt 60

gggttcgaat cccatcctcg tcg 83

<210> 429

<211> 75

<212> DNA

<213> Intelligent people

<400> 429

cggcgccgtg gcttagttgg ttaaagcgcc tgtctagtaa acaggagatc ctgggttcga 60

atcccagcgg tgcct 75

<210> 430

<211> 73

<212> DNA

<213> Intelligent people

<400> 430

ggcctcgtgg cgcaacggta gcgcgtctga ctccagatca gaaggttgcg tgttcaaatc 60

acgtcggggt caa 73

<210> 431

<211> 73

<212> DNA

<213> Intelligent people

<400> 431

agcgttggtg gtatagtggt aagcatagct gccttccaag cagttgaccc gggttcgatt 60

cccggccaac gca 73

<210> 432

<211> 73

<212> DNA

<213> Intelligent people

<400> 432

tcctcgttag tatagtggtg agtatccccg cctgtcacgc gggagaccgg ggttcgattc 60

cccgacgggg aga 73

<210> 433

<211> 73

<212> DNA

<213> Intelligent people

<400> 433

ggctcgttgg tctaggggta tgattctcgc ttcgggtgcg agaggtcccg ggttcaaatc 60

ccggacgagc cct 73

<210> 434

<211> 75

<212> DNA

<213> Intelligent people

<400> 434

ggcgccgtgg cttagttggt taaagcgcct gtctagtaaa caggagatcc tgggttcgaa 60

tcccagcggt gcctt 75

<210> 435

<211> 83

<212> DNA

<213> Intelligent people

<400> 435

gtagtcgtgg ccgagtggtt aaggcgatgg actagaaatc cattggggtc tccccgcgca 60

ggttcgaatc ctgccgacta cgt 83

<210> 436

<211> 73

<212> DNA

<213> Intelligent people

<400> 436

tgacctcgtg gcgcaatggt agcgcgtctg actccagatc agaaggttgc gtgttcaagt 60

cacgtcgggg tca 73

<210> 437

<211> 73

<212> DNA

<213> Intelligent people

<400> 437

aggcgcggtg gccaagtggt aaggcgtcgg tctcgtaaac cgaagatcgc gggttcgaac 60

cccgtccgtg cct 73

<210> 438

<211> 73

<212> DNA

<213> Intelligent people

<400> 438

agggggtata gctcagtggt agagcatttg actgcagatc aagaggtccc cggttcaaat 60

ccgggtgccc cct 73

<210> 439

<211> 73

<212> DNA

<213> Intelligent people

<400> 439

gggggtatag ctcagtggta gagcatttga ctgcagatca agaggtccct ggttcaaatc 60

cgggtgcccc ctc 73

<210> 440

<211> 73

<212> DNA

<213> Intelligent people

<400> 440

gggggtatag ctcagtggta gagcatttga ctgcagatca agaggtcccc ggttcaaatc 60

cgggtgcccc ctc 73

<210> 441

<211> 73

<212> DNA

<213> Intelligent people

<400> 441

aggtcccatg gtgtaatggt tagcactctg gactttgaat ccagcgatcc gagttcaaat 60

ctcggtggga cct 73

<210> 442

<211> 74

<212> DNA

<213> Intelligent people

<400> 442

gacccagtgg cctaatggat aaggcatcag cctccggagc tggggattgt gggttcgagt 60

cccatctggg tcgc 74

<210> 443

<211> 74

<212> DNA

<213> Intelligent people

<400> 443

agccccagtg gcctaatgga taaggcactg gcctcctaag ccagggattg tgggttcgag 60

tcccacctgg ggta 74

<210> 444

<211> 74

<212> DNA

<213> Intelligent people

<400> 444

gccccagtgg cctaatggat aaggcactgg cctcctaagc cagggattgt gggttcgagt 60

cccacctggg gtgt 74

<210> 445

<211> 74

<212> DNA

<213> Intelligent people

<400> 445

agaccgcgtg gcctaatgga taaggcgtct gacttcggat cagaagattg agggttcgag 60

tcccttcgtg gtcg 74

<210> 446

<211> 75

<212> DNA

<213> Intelligent people

<400> 446

cgtctctgtg gcgcaatcgg ttagcgcgtt cggctgttaa ccgaaaggtt ggtggttcga 60

gcccacccag ggacg 75

<210> 447

<211> 73

<212> DNA

<213> Intelligent people

<400> 447

ggcgttggtg gtatagtggt tagcatagct gccttccaag cagttgaccc gggttcgatt 60

cccggccaac gca 73

<210> 448

<211> 74

<212> DNA

<213> Intelligent people

<400> 448

gtttccgtag tgtagcggtt atcacattcg cctcacacgc gaaaggtccc cggttcgatc 60

ccgggcggaa acag 74

<210> 449

<211> 75

<212> DNA

<213> Intelligent people

<400> 449

tggcgccgtg gcttagttgg ttaaagcgcc tgtctagtaa acaggagatc ctgggttcga 60

atcccagcgg tgcct 75

<210> 450

<211> 94

<212> DNA

<213> Intelligent people

<400> 450

gctccagtgg cgcaatcggt tagcgcgcgg tacttatatg acagtgcgag cggagcaatg 60

ccgaggttgt gagttcgatc ctcacctgga gcac 94

<210> 451

<211> 72

<212> DNA

<213> Intelligent people

<400> 451

gcatgggtgg ttcagtggta gaattctcgc ctgccacgcg ggaggcccgg gttcgattcc 60

cggcccatgc ag 72

<210> 452

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 452

aaaatataaa tatatttc 18

<210> 453

<211> 5

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 453

aagct 5

<210> 454

<211> 5

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 454

aagtt 5

<210> 455

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 455

aattcttcgg aatgt 15

<210> 456

<211> 3

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 456

aga 3

<210> 457

<211> 5

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 457

agtcc 5

<210> 458

<211> 5

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 458

caacc 5

<210> 459

<211> 5

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 459

caatc 5

<210> 460

<211> 4

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 460

cagc 4

<210> 461

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 461

caggcgggtt ctgcccgcgc 20

<210> 462

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 462

catacctgca agggtatc 18

<210> 463

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 463

cgaccgcaag gttgt 15

<210> 464

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 464

cgaccttgcg gtcat 15

<210> 465

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 465

cgatgctaat cacatcgt 18

<210> 466

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 466

cgatggtgac atcat 15

<210> 467

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 467

cgatggttta catcgt 16

<210> 468

<211> 13

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 468

cgccgtaagg tgt 13

<210> 469

<211> 12

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 469

cgccttaggt gt 12

<210> 470

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 470

cgcctttcga cgcgt 15

<210> 471

<211> 13

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 471

cgcttcacgg cgt 13

<210> 472

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 472

cggcagcaat gctgt 15

<210> 473

<211> 13

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 473

cggctccgcc ttc 13

<210> 474

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 474

cgggtatcac agggtc 16

<210> 475

<211> 17

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 475

cggtgcgcaa gcgctgt 17

<210> 476

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 476

cgtacgggtg accgtacc 18

<210> 477

<211> 13

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 477

cgtcaaagac ttc 13

<210> 478

<211> 13

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 478

cgtcgtaaga ctt 13

<210> 479

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 479

cgttgaataa acgt 14

<210> 480

<211> 5

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 480

ctgtc 5

<210> 481

<211> 4

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 481

ggcc 4

<210> 482

<211> 7

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 482

ggggatt 7

<210> 483

<211> 4

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 483

ggtc 4

<210> 484

<211> 5

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 484

ggttt 5

<210> 485

<211> 4

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 485

gtag 4

<210> 486

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 486

taactagata ctttcagat 19

<210> 487

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 487

tactcgtatg ggtgc 15

<210> 488

<211> 13

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 488

tactttgcgg tgt 13

<210> 489

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 489

taggcgagta acatcgtgc 19

<210> 490

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 490

taggcgtgaa tagcgcctc 19

<210> 491

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 491

taggtcgcga gagcggcgc 19

<210> 492

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 492

taggtcgcgt aagcggcgc 19

<210> 493

<211> 17

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 493

taggtggtta tccacgc 17

<210> 494

<211> 5

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 494

tagtc 5

<210> 495

<211> 5

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 495

tagtt 5

<210> 496

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 496

tatacgtgaa agcgtatc 18

<210> 497

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 497

tatagggtca aaaactctat c 21

<210> 498

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 498

tatgcagaaa tacctgcatc 20

<210> 499

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 499

tccccatacg ggggc 15

<210> 500

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 500

tcccgaaggg gttc 14

<210> 501

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 501

tctacgtatg tgggc 15

<210> 502

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 502

tctcatagga gttc 14

<210> 503

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 503

tctcctctgg aggc 14

<210> 504

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 504

tcttagcaat aaggt 15

<210> 505

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 505

tcttgtagga gttc 14

<210> 506

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 506

tgaacgtaag ttcgc 15

<210> 507

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 507

tgaactgcga ggttcc 16

<210> 508

<211> 4

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 508

tgac 4

<210> 509

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 509

tgaccgaaag gtcgt 15

<210> 510

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 510

tgaccgcaag gtcgt 15

<210> 511

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 511

tgagctctgc tctc 14

<210> 512

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 512

tgaggcctca cggcctac 18

<210> 513

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 513

tgagggcaac ttcgt 15

<210> 514

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 514

tgagggtcat acctcc 16

<210> 515

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 515

tgagggtgca aatcctcc 18

<210> 516

<211> 13

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 516

tgccgaaagg cgt 13

<210> 517

<211> 13

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 517

tgccgtaagg cgt 13

<210> 518

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 518

tgcggtctcc gcgc 14

<210> 519

<211> 11

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 519

tgctagagca t 11

<210> 520

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 520

tgctcgtata gagctc 16

<210> 521

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 521

tggacaattg tctgc 15

<210> 522

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 522

tggacagatg tccgt 15

<210> 523

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 523

tggacaggtg tccgc 15

<210> 524

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 524

tggacggttg tccgc 15

<210> 525

<211> 13

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 525

tggacttgtg gtc 13

<210> 526

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 526

tggagattct ctccgc 16

<210> 527

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 527

tggcataggc ctgc 14

<210> 528

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 528

tggcttatgt ctac 14

<210> 529

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 529

tgggagttaa tcccgt 16

<210> 530

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 530

tgggatcttc ccgc 14

<210> 531

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 531

tgggcagaaa tgtctc 16

<210> 532

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 532

tgggcgttcg cccgc 15

<210> 533

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 533

tgggcttcgc ccgc 14

<210> 534

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 534

tgggggataa ccccgt 16

<210> 535

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 535

tgggggtttc cccgt 15

<210> 536

<211> 4

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 536

tggt 4

<210> 537

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 537

tggtggcaac accgt 15

<210> 538

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 538

tggtttatag ccgt 14

<210> 539

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 539

tgtacggtaa taccgtacc 19

<210> 540

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 540

tgtccgcaag gacgt 15

<210> 541

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 541

tgtcctaacg gacgt 15

<210> 542

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 542

tgtcctatta acggacgt 18

<210> 543

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 543

tgtccttcac gggcgt 16

<210> 544

<211> 13

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 544

tgtcttagga cgt 13

<210> 545

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 545

tgtgcgttaa cgcgtacc 18

<210> 546

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 546

tgtgtcgcaa ggcacc 16

<210> 547

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 547

tgttcgtaag gactt 15

<210> 548

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 548

ttcacagaaa tgtgtc 16

<210> 549

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 549

ttccctcgtg gagt 14

<210> 550

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 550

ttccctctgg gagc 14

<210> 551

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 551

ttcccttgtg gatc 14

<210> 552

<211> 13

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 552

ttccttcggg agc 13

<210> 553

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 553

ttctagcaat agagt 15

<210> 554

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 554

ttctccactg gggagc 16

<210> 555

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 555

ttctcgagag ggagc 15

<210> 556

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 556

ttctcgtatg agagc 15

<210> 557

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 557

tttaaggttt tcccttaac 19

<210> 558

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 558

tttcattgtg gagt 14

<210> 559

<211> 14

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 559

tttcgaagga atcc 14

<210> 560

<211> 13

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 560

tttcttcgga agc 13

<210> 561

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 561

tttggggcaa ctcaac 16

<210> 562

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (5)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (15)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (21)..(21)

<223> a, c, u, or g

<220>

<221> modified base

<222> (30)..(32)

<223> a, c, u, or g

<220>

<221> modified base

<222> (34)..(34)

<223> a, c, u, or g

<220>

<221> modified base

<222> (37)..(37)

<223> a, c, u, or g

<220>

<221> modified base

<222> (41)..(45)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (47)..(320)

<223> a, c, u, or g

<220>

<221> modified base

<222> (328)..(329)

<223> a, c, u, or g

<220>

<221> modified base

<222> (333)..(334)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(337)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 562

rkssnndurg hbyannyugr ndgwdvdydn nnbnhbnryr nnnnndnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnnnn rrduyranny ybnnhnnbwc cd 342

<210> 563

<211> 341

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (30)..(30)

<223> a, c, u, or g

<220>

<221> modified base

<222> (46)..(316)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (46)..(316)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (319)..(319)

<223> a, c, u, or g

<220>

<221> modified base

<222> (328)..(328)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(341)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(341)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 563

rksgrwdkrg hbyavnyggu dgarvrydyn hkywbhryrh dhdhrnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnbhnr gducrayncy ydvhhyywcc d 341

<210> 564

<211> 341

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (30)..(30)

<223> a, c, u, or g

<220>

<221> modified base

<222> (46)..(316)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (46)..(316)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (319)..(319)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(341)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(341)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 564

ggggrwdurg hbyavnyggu dgarvrydyn hkywkhryrh dhdhrnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnbhnr gducraybcc ydvhhyyucc a 341

<210> 565

<211> 343

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (2)..(8)

<223> a, c, u, or g

<220>

<221> modified base

<222> (12)..(13)

<223> a, c, u, or g

<220>

<221> modified base

<222> (17)..(17)

<223> a, c, u, or g

<220>

<221> modified base

<222> (22)..(24)

<223> a, c, u, or g

<220>

<221> modified base

<222> (26)..(27)

<223> a, c, u, or g

<220>

<221> modified base

<222> (30)..(35)

<223> a, c, u, or g

<220>

<221> modified base

<222> (38)..(38)

<223> a, c, u, or g

<220>

<221> modified base

<222> (43)..(43)

<223> a, c, u, or g

<220>

<221> modified base

<222> (45)..(322)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (48)..(318)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (329)..(329)

<223> a, c, u, or g

<220>

<221> modified base

<222> (333)..(342)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(343)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(343)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 565

gnnnnnnnkv gnnhddnhwr rnnnrnnvyn nnnnnbunck rhnbnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnnnn nnguuyrany ybnnnnnnnn nnd 343

<210> 566

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (3)..(3)

<223> a, c, u, or g

<220>

<221> modified base

<222> (6)..(7)

<223> a, c, u, or g

<220>

<221> modified base

<222> (13)..(13)

<223> a, c, u, or g

<220>

<221> modified base

<222> (26)..(26)

<223> a, c, u, or g

<220>

<221> modified base

<222> (32)..(32)

<223> a, c, u, or g

<220>

<221> modified base

<222> (37)..(37)

<223> a, c, u, or g

<220>

<221> modified base

<222> (44)..(44)

<223> a, c, u, or g

<220>

<221> modified base

<222> (47)..(317)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (328)..(328)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(338)

<223> a, c, u, or g

<220>

<221> modified base

<222> (340)..(340)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 566

grnvbnnvkb gbnydrwurg wygarnrydy ynsvyunckr mkbnrrnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnshv gguuyranuy cbdbynnnbn hr 342

<210> 567

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (3)..(3)

<223> a, c, u, or g

<220>

<221> modified base

<222> (7)..(7)

<223> a, c, u, or g

<220>

<221> modified base

<222> (47)..(317)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (336)..(336)

<223> a, c, u, or g

<220>

<221> modified base

<222> (340)..(340)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 567

grnvyvnskk gssydawugg aygarsgyry ykgmyuhckr akymrrnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnshr gguuygavuy cydbynbdbn yg 342

<210> 568

<211> 341

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (2)..(2)

<223> a, c, u, or g

<220>

<221> modified base

<222> (4)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (12)..(12)

<223> a, c, u, or g

<220>

<221> modified base

<222> (17)..(17)

<223> a, c, u, or g

<220>

<221> modified base

<222> (25)..(25)

<223> a, c, u, or g

<220>

<221> modified base

<222> (28)..(30)

<223> a, c, u, or g

<220>

<221> modified base

<222> (43)..(316)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (46)..(316)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (318)..(319)

<223> a, c, u, or g

<220>

<221> modified base

<222> (327)..(327)

<223> a, c, u, or g

<220>

<221> modified base

<222> (331)..(332)

<223> a, c, u, or g

<220>

<221> modified base

<222> (334)..(337)

<223> a, c, u, or g

<220>

<221> modified base

<222> (339)..(340)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(341)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(341)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 568

dnynnndurg hnhvrynggb hurdnrynnn bsryyruuwa hbnnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnvnnr gukhvwnhcy nnbnnnndnn r 341

<210> 569

<211> 341

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (17)..(17)

<223> a, c, u, or g

<220>

<221> modified base

<222> (28)..(28)

<223> a, c, u, or g

<220>

<221> modified base

<222> (46)..(316)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (46)..(316)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(341)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(341)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 569

ruyucygurg hryvrunggb yuarhgcnuu ysryyruuwa hbddannnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnrgur gukhvwdmcy ayysvrrrrb r 341

<210> 570

<211> 341

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (46)..(316)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (46)..(316)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(341)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(341)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 570

ruyucygurg hryvrubggb yuaghgcbuu ysryyruuaa hyddannnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnrgur gukhvwdmcy ayysvrrrry g 341

<210> 571

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (2)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (12)..(12)

<223> a, c, u, or g

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (22)..(22)

<223> a, c, u, or g

<220>

<221> modified base

<222> (26)..(26)

<223> a, c, u, or g

<220>

<221> modified base

<222> (29)..(29)

<223> a, c, u, or g

<220>

<221> modified base

<222> (31)..(32)

<223> a, c, u, or g

<220>

<221> modified base

<222> (44)..(44)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (47)..(319)

<223> a, c, u, or g

<220>

<221> modified base

<222> (328)..(328)

<223> a, c, u, or g

<220>

<221> modified base

<222> (333)..(334)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(339)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 571

knnnnndyrr ynhrrnyygr hnumdnrynb nnvhyurucr hdbnbvnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnnns rgyubrhnyy ysnnhnnnnb mr 342

<210> 572

<211> 339

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (2)..(2)

<223> a, c, u, or g

<220>

<221> modified base

<222> (6)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (44)..(314)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (44)..(314)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (333)..(333)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(339)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(339)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 572

knsbbnwyar ywyrgugguk mgwrubysyr yyurucaygb rsrnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnykgrgy ubrhkyycch rwnvvksmr 339

<210> 573

<211> 339

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (44)..(314)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (44)..(314)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(339)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(339)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 573

umcuyruyag uaurgugguk mguruyycyg ycugucaygy ggrnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnyggrgu uyrmkyyccy rasrrggag 339

<210> 574

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (3)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (12)..(13)

<223> a, c, u, or g

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (24)..(24)

<223> a, c, u, or g

<220>

<221> modified base

<222> (26)..(26)

<223> a, c, u, or g

<220>

<221> modified base

<222> (29)..(32)

<223> a, c, u, or g

<220>

<221> modified base

<222> (34)..(34)

<223> a, c, u, or g

<220>

<221> modified base

<222> (42)..(42)

<223> a, c, u, or g

<220>

<221> modified base

<222> (44)..(319)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (328)..(328)

<223> a, c, u, or g

<220>

<221> modified base

<222> (333)..(334)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(340)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 574

gsnnnndurv ynnavnkcgd byynrnvbnn nnvnyyrcar mnbnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnnnb druucranyy yvnnhnnnnn yy 342

<210> 575

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (45)..(315)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 575

gsgsryrkrg yubasnkgdk uaravyahuu grcyrcarau cmarnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnyyydr uucraruyyv gkuryychyy 340

<210> 576

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (45)..(315)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 576

gggsryrkrg yuyasnugrk uagagcayuu gacugcarau cmarnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnyyyrr uucraauyyv gkurcycmyy 340

<210> 577

<211> 341

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (2)..(2)

<223> a, c, u, or g

<220>

<221> modified base

<222> (4)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (12)..(13)

<223> a, c, u, or g

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (20)..(21)

<223> a, c, u, or g

<220>

<221> modified base

<222> (24)..(24)

<223> a, c, u, or g

<220>

<221> modified base

<222> (28)..(31)

<223> a, c, u, or g

<220>

<221> modified base

<222> (33)..(33)

<223> a, c, u, or g

<220>

<221> modified base

<222> (40)..(41)

<223> a, c, u, or g

<220>

<221> modified base

<222> (43)..(319)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (46)..(316)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (327)..(327)

<223> a, c, u, or g

<220>

<221> modified base

<222> (331)..(332)

<223> a, c, u, or g

<220>

<221> modified base

<222> (335)..(339)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(341)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(341)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 577

dnbnnnduvg bnnarnhggn nhanvvynnn nvnyubukrn nhnnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnnnr ruuyvanyby nnbhnnnnnh d 341

<210> 578

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (4)..(5)

<223> a, c, u, or g

<220>

<221> modified base

<222> (12)..(12)

<223> a, c, u, or g

<220>

<221> modified base

<222> (21)..(21)

<223> a, c, u, or g

<220>

<221> modified base

<222> (27)..(28)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (45)..(317)

<223> a, c, u, or g

<220>

<221> modified base

<222> (331)..(331)

<223> a, c, u, or g

<220>

<221> modified base

<222> (335)..(336)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 578

kdbnnydugg ynbarkmggd navvrynnhb vryubukrrb hvdvnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnndrr uuyvadybyh nbhdnnvhmd 340

<210> 579

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (4)..(4)

<223> a, c, u, or g

<220>

<221> modified base

<222> (21)..(21)

<223> a, c, u, or g

<220>

<221> modified base

<222> (27)..(28)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (45)..(317)

<223> a, c, u, or g

<220>

<221> modified base

<222> (331)..(331)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(336)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 579

kdbnvydugg ydbarumggk navvrynnws rryubukrab hsdvnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnndrr uuyvadysyh nbhdbnvhmk 340

<210> 580

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (1)..(2)

<223> a, c, u, or g

<220>

<221> modified base

<222> (5)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (9)..(9)

<223> a, c, u, or g

<220>

<221> modified base

<222> (12)..(12)

<223> a, c, u, or g

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (20)..(21)

<223> a, c, u, or g

<220>

<221> modified base

<222> (26)..(27)

<223> a, c, u, or g

<220>

<221> modified base

<222> (29)..(33)

<223> a, c, u, or g

<220>

<221> modified base

<222> (41)..(41)

<223> a, c, u, or g

<220>

<221> modified base

<222> (44)..(318)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (320)..(321)

<223> a, c, u, or g

<220>

<221> modified base

<222> (328)..(328)

<223> a, c, u, or g

<220>

<221> modified base

<222> (333)..(336)

<223> a, c, u, or g

<220>

<221> modified base

<222> (340)..(341)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 580

nnbbnndunr wnhrdnhygn nbhrdnnynn nnnvyuyusr nbvnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnnbn nryubrwnyy hbnnnndvvn nd 342

<210> 581

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (1)..(2)

<223> a, c, u, or g

<220>

<221> modified base

<222> (5)..(5)

<223> a, c, u, or g

<220>

<221> modified base

<222> (12)..(12)

<223> a, c, u, or g

<220>

<221> modified base

<222> (20)..(20)

<223> a, c, u, or g

<220>

<221> modified base

<222> (29)..(29)

<223> a, c, u, or g

<220>

<221> modified base

<222> (31)..(31)

<223> a, c, u, or g

<220>

<221> modified base

<222> (39)..(39)

<223> a, c, u, or g

<220>

<221> modified base

<222> (43)..(43)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (45)..(316)

<223> a, c, u, or g

<220>

<221> modified base

<222> (319)..(319)

<223> a, c, u, or g

<220>

<221> modified base

<222> (326)..(326)

<223> a, c, u, or g

<220>

<221> modified base

<222> (334)..(334)

<223> a, c, u, or g

<220>

<221> modified base

<222> (338)..(338)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 581

nnbynbdyrg unurdyagbn krrvvywbnb nvyuyusanb vhndnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnydnr yubrwnychb rkbndvvnvd 340

<210> 582

<211> 339

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (1)..(1)

<223> a, c, u, or g

<220>

<221> modified base

<222> (28)..(28)

<223> a, c, u, or g

<220>

<221> modified base

<222> (30)..(30)

<223> a, c, u, or g

<220>

<221> modified base

<222> (42)..(42)

<223> a, c, u, or g

<220>

<221> modified base

<222> (44)..(314)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (44)..(314)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (318)..(318)

<223> a, c, u, or g

<220>

<221> modified base

<222> (333)..(333)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(339)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(339)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 582

ncsydbrugg usuakygshk rrsaywbnkn syuyucahbv mndnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnbyrnry uyrwyucyyr gbnwrsraw 339

<210> 583

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (4)..(7)

<223> a, c, u, or g

<220>

<221> modified base

<222> (12)..(12)

<223> a, c, u, or g

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (21)..(22)

<223> a, c, u, or g

<220>

<221> modified base

<222> (26)..(26)

<223> a, c, u, or g

<220>

<221> modified base

<222> (29)..(34)

<223> a, c, u, or g

<220>

<221> modified base

<222> (41)..(319)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (328)..(328)

<223> a, c, u, or g

<220>

<221> modified base

<222> (333)..(338)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 583

rbvnnnnuvr ynyrrnyubd nnuadnrynn nnnnyybccv nnnnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnnnv vsubbrwnhc bbnnnnnnbb yw 342

<210> 584

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (5)..(5)

<223> a, c, u, or g

<220>

<221> modified base

<222> (28)..(28)

<223> a, c, u, or g

<220>

<221> modified base

<222> (39)..(40)

<223> a, c, u, or g

<220>

<221> modified base

<222> (42)..(42)

<223> a, c, u, or g

<220>

<221> modified base

<222> (45)..(315)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (317)..(317)

<223> a, c, u, or g

<220>

<221> modified base

<222> (335)..(335)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 584

gbvbnsvurr udyrrdcggk dadmaudnhh rhyubccann ynkdnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnbnrgs uubrwuuccy ksbsnvygca 340

<210> 585

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (5)..(5)

<223> a, c, u, or g

<220>

<221> modified base

<222> (28)..(28)

<223> a, c, u, or g

<220>

<221> modified base

<222> (39)..(39)

<223> a, c, u, or g

<220>

<221> modified base

<222> (42)..(42)

<223> a, c, u, or g

<220>

<221> modified base

<222> (45)..(315)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (335)..(335)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 585

gyvynsvurr udyrrwcggk kadmaudnhh rhyubccand ynkdnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnbvrgs uuyrwuuccy ksbsnvygca 340

<210> 586

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (4)..(7)

<223> a, c, u, or g

<220>

<221> modified base

<222> (13)..(13)

<223> a, c, u, or g

<220>

<221> modified base

<222> (25)..(26)

<223> a, c, u, or g

<220>

<221> modified base

<222> (29)..(31)

<223> a, c, u, or g

<220>

<221> modified base

<222> (34)..(34)

<223> a, c, u, or g

<220>

<221> modified base

<222> (42)..(42)

<223> a, c, u, or g

<220>

<221> modified base

<222> (45)..(317)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (319)..(320)

<223> a, c, u, or g

<220>

<221> modified base

<222> (328)..(328)

<223> a, c, u, or g

<220>

<221> modified base

<222> (332)..(333)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(338)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 586

kgynnnnduv gbnhagbyug ghyannvynn ndbnyugugr hnvhnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnvnn rgyucranyc ynnbhnnnvr ym 342

<210> 587

<211> 339

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (44)..(314)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (44)..(314)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(339)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(339)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 587

gchrugaybg uayagkgguu asyacucugy gyuguggccr cagnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnncwyggy ucraaucyga gucaygrca 339

<210> 588

<211> 339

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (44)..(314)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (44)..(314)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(339)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(339)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 588

gcmrugayyg uauagugguu agyacucugy gyuguggccr cagnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnncwyggu ucraaucyga gucaygrca 339

<210> 589

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (1)..(1)

<223> a, c, u, or g

<220>

<221> modified base

<222> (4)..(5)

<223> a, c, u, or g

<220>

<221> modified base

<222> (7)..(7)

<223> a, c, u, or g

<220>

<221> modified base

<222> (17)..(17)

<223> a, c, u, or g

<220>

<221> modified base

<222> (31)..(32)

<223> a, c, u, or g

<220>

<221> modified base

<222> (34)..(34)

<223> a, c, u, or g

<220>

<221> modified base

<222> (37)..(37)

<223> a, c, u, or g

<220>

<221> modified base

<222> (42)..(320)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (328)..(329)

<223> a, c, u, or g

<220>

<221> modified base

<222> (332)..(334)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(339)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 589

nvsnnbndur gybyrrnywg gbhrdvrydb nnbnyunaur annnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnnnn dsywydannc hnnnhnnnns ba 342

<210> 590

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (45)..(315)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (326)..(327)

<223> a, c, u, or g

<220>

<221> modified base

<222> (335)..(335)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 590

gsyybrkurg ykyrruyggy hagmgcrygk urcudauaay ryyrnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnryrrs ywydanncyc rymyngrsca 340

<210> 591

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (45)..(315)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 591

gsyysrkurg ykcaruyggy hagmgcgygg urcudauaay rcyrnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnngygrg yucrabhcyc rymybgrsca 340

<210> 592

<211> 341

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (1)..(1)

<223> a, c, u, or g

<220>

<221> modified base

<222> (3)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (11)..(12)

<223> a, c, u, or g

<220>

<221> modified base

<222> (16)..(17)

<223> a, c, u, or g

<220>

<221> modified base

<222> (21)..(22)

<223> a, c, u, or g

<220>

<221> modified base

<222> (25)..(26)

<223> a, c, u, or g

<220>

<221> modified base

<222> (28)..(31)

<223> a, c, u, or g

<220>

<221> modified base

<222> (41)..(41)

<223> a, c, u, or g

<220>

<221> modified base

<222> (43)..(316)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (46)..(316)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (318)..(319)

<223> a, c, u, or g

<220>

<221> modified base

<222> (327)..(327)

<223> a, c, u, or g

<220>

<221> modified base

<222> (331)..(332)

<223> a, c, u, or g

<220>

<221> modified base

<222> (335)..(338)

<223> a, c, u, or g

<220>

<221> modified base

<222> (340)..(340)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(341)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(341)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 592

nbnnnndurs nnbarnnhgg nnddnnbnnn nvdhycauah nbnnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnvnnr gdusdanhmy nnbhnnnnvn w 341

<210> 593

<211> 339

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (4)..(4)

<223> a, c, u, or g

<220>

<221> modified base

<222> (44)..(314)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (44)..(314)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(339)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(339)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 593

rbcnbvkurg ygcagydggh agyryryyrg kyycauaahy yvrnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnsdkrgd usdadmmymw smbvbgsya 339

<210> 594

<211> 339

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (44)..(314)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (44)..(314)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(339)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(339)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 594

rschbvkurg ygcagydggh agcryryyrg kyycauaayc yrrnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnswkrgw usdadmcymw smbvkgsya 339

<210> 595

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (2)..(7)

<223> a, c, u, or g

<220>

<221> modified base

<222> (9)..(12)

<223> a, c, u, or g

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (23)..(23)

<223> a, c, u, or g

<220>

<221> modified base

<222> (26)..(26)

<223> a, c, u, or g

<220>

<221> modified base

<222> (28)..(34)

<223> a, c, u, or g

<220>

<221> modified base

<222> (37)..(37)

<223> a, c, u, or g

<220>

<221> modified base

<222> (41)..(317)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (319)..(320)

<223> a, c, u, or g

<220>

<221> modified base

<222> (328)..(328)

<223> a, c, u, or g

<220>

<221> modified base

<222> (332)..(333)

<223> a, c, u, or g

<220>

<221> modified base

<222> (335)..(340)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 595

vnnnnnnunn nnvrrnhhgk hhndhnvnnn nnnnhynard nnnnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnvnn vbyuhvanym bnnbnnnnnn br 342

<210> 596

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (6)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (33)..(33)

<223> a, c, u, or g

<220>

<221> modified base

<222> (41)..(41)

<223> a, c, u, or g

<220>

<221> modified base

<222> (43)..(43)

<223> a, c, u, or g

<220>

<221> modified base

<222> (46)..(317)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (319)..(319)

<223> a, c, u, or g

<220>

<221> modified base

<222> (328)..(328)

<223> a, c, u, or g

<220>

<221> modified base

<222> (333)..(333)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(336)

<223> a, c, u, or g

<220>

<221> modified base

<222> (340)..(340)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 596

rbbvvndurk hhsrrbhygk hhurwbrhdb hdnryuhard nynhdnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnvnv gbyuhvanym ybnbbnbbvn ya 342

<210> 597

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (6)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (33)..(33)

<223> a, c, u, or g

<220>

<221> modified base

<222> (43)..(43)

<223> a, c, u, or g

<220>

<221> modified base

<222> (46)..(317)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (319)..(319)

<223> a, c, u, or g

<220>

<221> modified base

<222> (333)..(333)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(336)

<223> a, c, u, or g

<220>

<221> modified base

<222> (340)..(340)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 597

rbbvvndurk hhsrrbhygk hhurwkrhdb hdnryuhard hynhdnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnvnv gbyuhvavym ybnbynbbvn ya 342

<210> 598

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (1)..(7)

<223> a, c, u, or g

<220>

<221> modified base

<222> (11)..(12)

<223> a, c, u, or g

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (21)..(21)

<223> a, c, u, or g

<220>

<221> modified base

<222> (26)..(26)

<223> a, c, u, or g

<220>

<221> modified base

<222> (30)..(32)

<223> a, c, u, or g

<220>

<221> modified base

<222> (44)..(320)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (328)..(328)

<223> a, c, u, or g

<220>

<221> modified base

<222> (332)..(333)

<223> a, c, u, or g

<220>

<221> modified base

<222> (335)..(340)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 598

nnnnnnnurs nnharnhwrr nuudrnrydn nnsvyyyuum mbvnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnnnn vryuyrwnhy bnnbnnnnnn hd 342

<210> 599

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (1)..(1)

<223> a, c, u, or g

<220>

<221> modified base

<222> (6)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (11)..(11)

<223> a, c, u, or g

<220>

<221> modified base

<222> (29)..(29)

<223> a, c, u, or g

<220>

<221> modified base

<222> (43)..(43)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (45)..(317)

<223> a, c, u, or g

<220>

<221> modified base

<222> (331)..(331)

<223> a, c, u, or g

<220>

<221> modified base

<222> (333)..(334)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(336)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 599

nhvbdnvkrs nhharbhrrb udrdrydbnd gryyyuummy mhndnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnrgr yuyrwbyysy nbnnhndrhr 340

<210> 600

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (45)..(316)

<223> a, c, u, or g

<220>

<221> modified base

<222> (333)..(333)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(336)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 600

vhvbrdvkas cwharuhrrb udrwrcrkvd gacuyuumau chswnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnndrgr yuyrwkyysy hbnhynkryr 340

<210> 601

<211> 341

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (4)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (29)..(31)

<223> a, c, u, or g

<220>

<221> modified base

<222> (33)..(33)

<223> a, c, u, or g

<220>

<221> modified base

<222> (41)..(41)

<223> a, c, u, or g

<220>

<221> modified base

<222> (43)..(318)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (46)..(316)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (327)..(327)

<223> a, c, u, or g

<220>

<221> modified base

<222> (331)..(332)

<223> a, c, u, or g

<220>

<221> modified base

<222> (335)..(338)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(341)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(341)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 601

kbbnnnduag yyyarhyugg bwwgrryrnn nbnyygaarv nbnnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnnbd guucranych nnbhnnnnvm a 341

<210> 602

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (45)..(315)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 602

kbyvwrruwg yyyaruuggs uagrrydykr brcygaarry syhmnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnyckgg uucrayycys gguywbrvma 340

<210> 603

<211> 339

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (44)..(314)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (44)..(314)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(339)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(339)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 603

gcyrarauwg cucaruuggg agagyguuas acygaagauc uwmnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnycuggu ucrayycygg guuucrvca 339

<210> 604

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (4)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (21)..(21)

<223> a, c, u, or g

<220>

<221> modified base

<222> (29)..(34)

<223> a, c, u, or g

<220>

<221> modified base

<222> (37)..(37)

<223> a, c, u, or g

<220>

<221> modified base

<222> (41)..(320)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (328)..(328)

<223> a, c, u, or g

<220>

<221> modified base

<222> (331)..(334)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(338)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 604

sksnnnduvg bbyagnyygb nyyrdvvynn nnnnhunggr nnnnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnnnn dguucranyc nnnnhnnnbm sm 342

<210> 605

<211> 338

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (39)..(39)

<223> a, c, u, or g

<220>

<221> modified base

<222> (43)..(313)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (43)..(313)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(338)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(338)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 605

ggcusguugg kcuagkgbur ugruucucrs yuhggrysnr agnnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnncyggguu caaaucvyrg asgagccc 338

<210> 606

<211> 338

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (43)..(313)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (43)..(313)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(338)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(338)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 606

ggcucguugg ucuagkggur ugruucucgs uuhggrysbg agnnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnccggguu caaaucccgg acgagccc 338

<210> 607

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (2)..(7)

<223> a, c, u, or g

<220>

<221> modified base

<222> (9)..(13)

<223> a, c, u, or g

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (21)..(21)

<223> a, c, u, or g

<220>

<221> modified base

<222> (25)..(28)

<223> a, c, u, or g

<220>

<221> modified base

<222> (30)..(34)

<223> a, c, u, or g

<220>

<221> modified base

<222> (37)..(37)

<223> a, c, u, or g

<220>

<221> modified base

<222> (42)..(320)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (332)..(340)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 607

dnnnnnnynn nnnarnbhgg nbbannnndn nnnnyynswr mnnnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnnnn rkuuyradyc ynnnnnnnnn bd 342

<210> 608

<211> 341

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (2)..(5)

<223> a, c, u, or g

<220>

<221> modified base

<222> (36)..(36)

<223> a, c, u, or g

<220>

<221> modified base

<222> (46)..(316)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (46)..(316)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (319)..(319)

<223> a, c, u, or g

<220>

<221> modified base

<222> (331)..(331)

<223> a, c, u, or g

<220>

<221> modified base

<222> (335)..(336)

<223> a, c, u, or g

<220>

<221> modified base

<222> (338)..(339)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(341)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(341)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 608

knnnnbdurg chsarkbugg uuavdghrdb kdrcynswra ybmvdnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnvyng kuuyradycc nrbhnnbnny d 341

<210> 609

<211> 341

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (2)..(3)

<223> a, c, u, or g

<220>

<221> modified base

<222> (5)..(5)

<223> a, c, u, or g

<220>

<221> modified base

<222> (36)..(36)

<223> a, c, u, or g

<220>

<221> modified base

<222> (46)..(316)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (46)..(316)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (335)..(336)

<223> a, c, u, or g

<220>

<221> modified base

<222> (338)..(339)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(341)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(341)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 609

knndnbrurg chsarkcugg uuavdghrwy krrcunswra yymrdnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnvydg guuyraaycc hrbynnynny k 341

<210> 610

<211> 341

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (4)..(7)

<223> a, c, u, or g

<220>

<221> modified base

<222> (12)..(12)

<223> a, c, u, or g

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (25)..(25)

<223> a, c, u, or g

<220>

<221> modified base

<222> (29)..(31)

<223> a, c, u, or g

<220>

<221> modified base

<222> (33)..(33)

<223> a, c, u, or g

<220>

<221> modified base

<222> (36)..(36)

<223> a, c, u, or g

<220>

<221> modified base

<222> (41)..(41)

<223> a, c, u, or g

<220>

<221> modified base

<222> (43)..(319)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (46)..(316)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (327)..(327)

<223> a, c, u, or g

<220>

<221> modified base

<222> (331)..(337)

<223> a, c, u, or g

<220>

<221> modified base

<222> (341)..(341)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(341)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(341)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 610

rbbnnnnurg ynhadnhykg hyrdnryrnn nsnhynguwa nsnnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnnnd gwyyvanbyh nnnnnnnrvy n 341

<210> 611

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (4)..(4)

<223> a, c, u, or g

<220>

<221> modified base

<222> (6)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (12)..(12)

<223> a, c, u, or g

<220>

<221> modified base

<222> (24)..(24)

<223> a, c, u, or g

<220>

<221> modified base

<222> (45)..(315)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (317)..(317)

<223> a, c, u, or g

<220>

<221> modified base

<222> (326)..(326)

<223> a, c, u, or g

<220>

<221> modified base

<222> (331)..(332)

<223> a, c, u, or g

<220>

<221> modified base

<222> (334)..(334)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(336)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 611

rbbnbnduvg ynharbyggh yrdnryryhb sbcyhguwav svdrnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnvndrg wuyvanbyyh nnbndnrvyb 340

<210> 612

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (4)..(4)

<223> a, c, u, or g

<220>

<221> modified base

<222> (6)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (12)..(12)

<223> a, c, u, or g

<220>

<221> modified base

<222> (24)..(24)

<223> a, c, u, or g

<220>

<221> modified base

<222> (45)..(315)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (326)..(326)

<223> a, c, u, or g

<220>

<221> modified base

<222> (332)..(332)

<223> a, c, u, or g

<220>

<221> modified base

<222> (334)..(334)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(336)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 612

rsynbnduvg ynharbuggh yrdnryryhk sbcyhguaav smdrnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnvhdrg wuyvanbyyh dnbndnrsyu 340

<210> 613

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (4)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (29)..(32)

<223> a, c, u, or g

<220>

<221> modified base

<222> (34)..(34)

<223> a, c, u, or g

<220>

<221> modified base

<222> (42)..(42)

<223> a, c, u, or g

<220>

<221> modified base

<222> (44)..(317)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (319)..(319)

<223> a, c, u, or g

<220>

<221> modified base

<222> (321)..(321)

<223> a, c, u, or g

<220>

<221> modified base

<222> (328)..(328)

<223> a, c, u, or g

<220>

<221> modified base

<222> (333)..(333)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(338)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 613

rvvnnndurr ybhhrhyhgg hyuadvvynn nnrnbuccav anbnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnvnr nguucranyc hynbhnnnbb hd 342

<210> 614

<211> 339

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (44)..(314)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (44)..(314)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(339)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(339)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 614

grmmkvrkgg ysmaryuggh arssykkybr rsuccasakb vrwnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnsygkgu ucradycmcr kyksbgyma 339

<210> 615

<211> 339

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (44)..(314)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (44)..(314)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(339)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(339)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 615

grmmkvrkgg ysmaryuggh arssykkybr rsuccasakb vrwnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnsygkgu ucradycmcr kyksbgyma 339

<210> 616

<211> 342

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (3)..(6)

<223> a, c, u, or g

<220>

<221> modified base

<222> (9)..(10)

<223> a, c, u, or g

<220>

<221> modified base

<222> (12)..(13)

<223> a, c, u, or g

<220>

<221> modified base

<222> (16)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (25)..(26)

<223> a, c, u, or g

<220>

<221> modified base

<222> (30)..(32)

<223> a, c, u, or g

<220>

<221> modified base

<222> (42)..(42)

<223> a, c, u, or g

<220>

<221> modified base

<222> (44)..(317)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (47)..(317)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (319)..(320)

<223> a, c, u, or g

<220>

<221> modified base

<222> (328)..(328)

<223> a, c, u, or g

<220>

<221> modified base

<222> (332)..(333)

<223> a, c, u, or g

<220>

<221> modified base

<222> (336)..(340)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(342)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(342)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 616

bbnnnndunn ynnarnyugs yhwrnnrbdn nnrdcuruar vnynnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnvnn vgwuyranuc bnnbhnnnnn vd 342

<210> 617

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (45)..(315)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (336)..(336)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 617

bbkkbdauag yucagbugsy uagagydkwk racuruagrk ymwknnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnscugg wuyrahucyr rsubsnmsvd 340

<210> 618

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (45)..(315)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 618

bbkksdauag yucagyuggy uagagcdkwk gacuruagrk ymwknnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnscugg uuyrawuccr rsuysdmsva 340

<210> 619

<211> 341

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (2)..(2)

<223> a, c, u, or g

<220>

<221> modified base

<222> (4)..(7)

<223> a, c, u, or g

<220>

<221> modified base

<222> (12)..(12)

<223> a, c, u, or g

<220>

<221> modified base

<222> (15)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (21)..(21)

<223> a, c, u, or g

<220>

<221> modified base

<222> (24)..(25)

<223> a, c, u, or g

<220>

<221> modified base

<222> (28)..(28)

<223> a, c, u, or g

<220>

<221> modified base

<222> (30)..(33)

<223> a, c, u, or g

<220>

<221> modified base

<222> (36)..(36)

<223> a, c, u, or g

<220>

<221> modified base

<222> (40)..(319)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (46)..(316)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (327)..(327)

<223> a, c, u, or g

<220>

<221> modified base

<222> (330)..(339)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(341)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(341)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 619

dndnnnnuvr ynbrnnhugk nyannrynbn nnnhunacrn nnnnnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnnnnnd rdubranhyn nnnnnnnnny v 341

<210> 620

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (15)..(16)

<223> a, c, u, or g

<220>

<221> modified base

<222> (24)..(24)

<223> a, c, u, or g

<220>

<221> modified base

<222> (27)..(27)

<223> a, c, u, or g

<220>

<221> modified base

<222> (29)..(30)

<223> a, c, u, or g

<220>

<221> modified base

<222> (41)..(43)

<223> a, c, u, or g

<220>

<221> modified base

<222> (45)..(315)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (317)..(318)

<223> a, c, u, or g

<220>

<221> modified base

<222> (326)..(326)

<223> a, c, u, or g

<220>

<221> modified base

<222> (330)..(330)

<223> a, c, u, or g

<220>

<221> modified base

<222> (333)..(333)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 620

kbkbshdurr ydbrnnhgku yadnrynynn dhyuhacrhk nnnrnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnbnnrr dubranhybn dvndsdwvyv 340

<210> 621

<211> 340

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Polynucleotide "

<220>

<221> modified base

<222> (45)..(315)

<223> a, c, u, or g

<220>

<221> features not yet classified

<222> (45)..(315)

<223 >/Note = "the region may cover 1-271 nucleotides"

<220>

<221> modified base

<222> (317)..(318)

<223> a, c, u, or g

<220>

<221> modified base

<222> (330)..(330)

<223> a, c, u, or g

<220>

<221> variants

<222> (1)..(340)

<223 >/replace = ""

<220>

<221> features not yet classified

<222> (1)..(340)

<223 >/Note = "No preference given to variant residues in sequence with respect to residues in the variant position Note"

<220>

<221> sources

<223 >/Note = "detailed description on substitution and preferred embodiment, please refer to the filed specification"

<400> 621

kbkbsydurg ydbrbhugku yadhryvyhh dyyuhacahk hddrnnnnnn nnnnnnnnnn 60

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 120

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 180

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 240

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 300

nnnnnnnnnn nnnnnbnngr dubradhyyn dvhrsdwvya 340

<210> 622

<211> 72

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 622

agcagagtgg cgcagcggaa gcgtgctggg cccataaccc agaggtcgat ggatcgaaac 60

catcctctgc ta 72

<210> 623

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Primers "

<400> 623

tgagttggca acctgtggta 20

<210> 624

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Primers "

<400> 624

ttgggtgtcc atgaaaatca 20

<210> 625

<211> 26

<212> RNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Probe'

<400> 625

uagcagagga ugguuucgau ccauca 26

<210> 626

<211> 72

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Oligonucleotides "

<400> 626

agcagagtgg cgcagcggaa gcgtgctggg cccataaccc agaggtcgat ggatcgaaac 60

catcctctgc ta 72

<210> 627

<211> 26

<212> DNA

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

Probe'

<400> 627

tagcagagga tggtttcgat ccatca 26

<210> 628

<211> 6

<212> PRT

<213> Artificial sequence

<220>

<221> sources

<223 >/Note = "description of Artificial sequence-synthetic

6XHis tag "

<400> 628

His His His His His His

1 5

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