阅读说明：本技术 生物电子器件及其制备方法和可控转化方法 (Bioelectronic device, method for producing the same, and controllable conversion method ) 是由 孙伟 陈雅鸿 于 2020-05-14 设计创作，主要内容包括：本公开提供了一种生物电子器件,包括：基片；第一电极,第一电极位于基片上；第二电极,第二电极位于基片上,并且与第一电极间隔设置；以及自组装核酸超结构,自组装核酸超结构位于第一电极和第二电极之间,其中自组装核酸超结构由特定数量、特定序列和特定取向的单链核酸构筑,并且通过施加至第一电极和第二电极之间的电场的调控,来实现双链核酸与单链核酸在特定数量、特定序列和特定取向上的可控转化。本公开还提供了一种生物电子器件的制备方法及可控转化方法。(The present disclosure provides a bioelectronic device comprising: a substrate; a first electrode on the substrate; a second electrode on the substrate and spaced apart from the first electrode; and a self-assembled nucleic acid superstructure located between the first electrode and the second electrode, wherein the self-assembled nucleic acid superstructure is constructed from a specific amount, a specific sequence and a specific orientation of single-stranded nucleic acids, and controlled transformation of double-stranded nucleic acids with single-stranded nucleic acids in the specific amount, specific sequence and specific orientation is achieved by regulation of an electric field applied between the first electrode and the second electrode. The disclosure also provides a preparation method and a controllable conversion method of the bioelectronic device.)

1. A bioelectronic device, comprising:

a substrate;

a first electrode on the substrate;

a second electrode on the substrate and spaced apart from the first electrode; and

a self-assembled nucleic acid superstructure located between the first electrode and the second electrode,

wherein the self-assembled nucleic acid superstructure is constructed from a specific number, a specific sequence and a specific orientation of single-stranded nucleic acids, and controlled conversion of the specific number, specific sequence and specific orientation of double-stranded nucleic acids to single-stranded nucleic acids is achieved by modulation of the electric field applied between the first electrode and the second electrode.

2. The bioelectronic device according to claim 1, wherein said first electrode and said second electrode are applied with different voltages for a controlled conversion of a specific amount, a specific sequence and a specific orientation of double stranded nucleic acids and single stranded nucleic acids.

3. A method of fabricating a bioelectronic device, comprising:

synthesizing a self-assembled nucleic acid superstructure constructed from a specific number, a specific sequence and a specific orientation of single-stranded nucleic acids;

depositing a self-assembled nucleic acid superstructure constructed from a specific number, a specific sequence and a specific orientation of single stranded nucleic acids on a substrate; and

processing a first electrode and a second electrode at predetermined locations of a surface of a deposited self-assembled nucleic acid superstructure, wherein the self-assembled nucleic acid superstructure is located between the first and second electrodes,

wherein the controllable conversion of a specific amount, a specific sequence and a specific orientation of double stranded nucleic acids to single stranded nucleic acids is achieved by the regulation of the electric field applied between the first and second electrodes.

4. The method of claim 3, wherein the first electrode and the second electrode are capable of being applied with different voltages to achieve a controlled conversion of a specific amount, a specific sequence and a specific orientation of double stranded nucleic acids to single stranded nucleic acids.

5. The method of claim 3 or 4, wherein processing the first electrode and the second electrode at predetermined locations on the surface of the deposited self-assembled nucleic acid superstructure comprises:

coating a photoresist layer on the surface of the self-assembled nucleic acid superstructure;

writing a first electrode and a second electrode pattern;

developing the patterns of the first and second electrodes by a developing solution; and

and stripping the photoresist layer by a stripping liquid after the metal film is deposited.

6. The method of any of claims 3 to 5, wherein depositing a self-assembled nucleic acid superstructure constructed from a specific number, a specific sequence and a specific orientation of single stranded nucleic acids onto a substrate comprises:

incubating for a predetermined time after depositing the self-assembled nucleic acid superstructure on the surface of the substrate;

after removing the remaining solution, performing a desalting process to remove residual inorganic salts from the substrate; and

and after drying, carrying out imaging treatment, and imaging the self-assembled nucleic acid superstructure on the substrate.

7. A method for the controlled conversion of double stranded nucleic acids to single stranded nucleic acids by the bioelectronic device according to claim 1 or 2 or by the bioelectronic device prepared by the method according to any one of claims 3 to 6,

the electric field regulation is carried out through the first electrode and the second electrode, and the controllable transformation of specific quantity, specific sequence and specific orientation of double-stranded nucleic acid and single-stranded nucleic acid is realized.

8. A system for selective control of protein binding, comprising: the bioelectronic device according to claim 1 or 2 or prepared by the method according to any one of claims 3 to 6,

wherein the single-stranded nucleic acid is designed to have a specific aptamer or to comprise a protein binding site, and the aptamer sequence or the protein binding sequence is controllably exposed when an electric field is applied to the first electrode and the second electrode of the bioelectronic device, thereby controllably capturing the protein.

9. A nanomaterial-bonded selective control system comprising: the bioelectronic device according to claim 1 or 2 or prepared by the method according to any one of claims 3 to 6,

wherein the surface of the nanomaterial is modified by the single-stranded nucleic acid handle, the single-stranded nucleic acid counter handle is introduced into the bioelectronic device, and when an electric field is applied to the first electrode and the second electrode of the bioelectronic device, the single-stranded nucleic acid counter handle is controllably exposed, and the nanomaterial modified by the single-stranded nucleic acid handle is controllably bound to the bioelectronic device.

10. A combining circuit, comprising: the bioelectronic device according to claim 1 or 2 or prepared by the method according to any one of claims 3 to 6,

wherein nucleic acid sequences binding to different disease markers are provided to the bioelectronic device, the binding sequences are selectively converted when an electric field is applied to the first and second electrodes of the bioelectronic device, the converted binding sequences bind to the disease markers in the synthetic circuit, and a biomolecular output signal is generated to trigger the synthetic circuit, thereby obtaining a diagnostic result of the disease.

Technical Field

The disclosure relates to the field of bioelectronics, in particular to a bioelectronics device and a preparation method and a controllable conversion method thereof.

Background

In the prior art, the controllable transformation of single-stranded nucleic acid and double-stranded nucleic acid can be controlled by means of light, chemical reagents and the like. For example, on a solid surface, a specific nucleic acid strand can be selectively cleaved by introducing a photocleavable nucleotide, resulting in conversion of a double-stranded nucleic acid into a single-stranded nucleic acid. Similarly, single-stranded nucleic acids at specific locations can be modified by super-resolution microscopy to achieve quantitative, sequence-specific, and orientation-specific controllable transformation.

However, in the field of electrically controlled switches, it is still difficult to achieve quantitative, sequencing and spatially oriented controlled conversion of single-stranded and double-stranded nucleic acids. Although the prior art realizes the modification of single-stranded nucleic acid on the surfaces of carbon nanotubes, silicon nanowires and two-dimensional materials. However, in these techniques, due to the random distribution characteristic of the modification chemical reaction itself, the single-stranded nucleic acids are distributed on the surface of the electronic material uncontrollably, and thus the number and relative spatial positions of the single-stranded nucleic acids cannot be controlled.

Disclosure of Invention

To address at least one of the above technical problems, the present disclosure provides a bioelectronic device, a method of manufacturing a bioelectronic device, and a controlled conversion method.

According to one aspect of the present disclosure, a bioelectronic device includes:

a substrate;

a first electrode on the substrate;

a second electrode on the substrate and spaced apart from the first electrode; and

a self-assembled nucleic acid superstructure located between the first electrode and the second electrode,

wherein the self-assembled nucleic acid superstructure is constructed from a specific number, a specific sequence and a specific orientation of single-stranded nucleic acids, and controlled conversion of the specific number, specific sequence and specific orientation of double-stranded nucleic acids to single-stranded nucleic acids is achieved by modulation of the electric field applied between the first electrode and the second electrode.

According to at least one embodiment of the present disclosure, the first electrode and the second electrode are applied with different voltages, enabling a controlled conversion of a specific amount, a specific sequence and a specific orientation of double stranded nucleic acids and single stranded nucleic acids.

According to another aspect of the present disclosure, a method of fabricating a bioelectronic device includes:

synthesizing a self-assembled nucleic acid superstructure constructed from a specific number, a specific sequence and a specific orientation of single-stranded nucleic acids;

depositing a self-assembled nucleic acid superstructure constructed from a specific number, a specific sequence and a specific orientation of single stranded nucleic acids on a substrate; and

processing a first electrode and a second electrode at predetermined locations of a surface of a deposited self-assembled nucleic acid superstructure, wherein the self-assembled nucleic acid superstructure is located between the first and second electrodes,

wherein the controllable conversion of a specific amount, a specific sequence and a specific orientation of double stranded nucleic acids to single stranded nucleic acids is achieved by the regulation of the electric field applied between the first and second electrodes.

According to at least one embodiment of the present disclosure, the first electrode and the second electrode are capable of being applied with different voltages, enabling a controlled conversion of a specific amount, a specific sequence and a specific orientation of double stranded nucleic acids to single stranded nucleic acids.

According to at least one embodiment of the present disclosure, processing the first electrode and the second electrode at predetermined locations on the surface of the self-assembled nucleic acid superstructure after deposition comprises:

coating a photoresist layer on the surface of the self-assembled nucleic acid superstructure;

writing a first electrode and a second electrode pattern;

developing the patterns of the first and second electrodes by a developing solution; and

and stripping the photoresist layer by a stripping liquid after the metal film is deposited.

According to at least one embodiment of the present disclosure, depositing a self-assembled nucleic acid superstructure constructed from a specific number, a specific sequence and a specific orientation of single-stranded nucleic acids on a substrate comprises:

incubating for a predetermined time after depositing the self-assembled nucleic acid superstructure on the surface of the substrate;

after removing the remaining solution, performing a desalting process to remove residual inorganic salts from the substrate; and

and after drying, carrying out imaging treatment, and imaging the self-assembled nucleic acid superstructure on the substrate.

According to yet another aspect of the present disclosure, a method for performing controlled conversion of double-stranded nucleic acids and single-stranded nucleic acids by the bioelectronic device as described above or by the bioelectronic device prepared by the method as described above, wherein the electric field is controlled by the first electrode and the second electrode, thereby achieving controlled conversion of the double-stranded nucleic acids and the single-stranded nucleic acids in a specific amount, a specific sequence and a specific orientation.

According to yet another aspect of the present disclosure, a protein binding selection control system includes: the bioelectronic device as described above or the bioelectronic device prepared by the method as described above,

wherein the single-stranded nucleic acid is designed to have a specific aptamer or to comprise a protein binding site, and the aptamer sequence or the protein binding sequence is controllably exposed when an electric field is applied to the first electrode and the second electrode of the bioelectronic device, thereby controllably capturing the protein.

According to yet another aspect of the present disclosure, a nanomaterial-bound selection control system includes: the bioelectronic device as described above or the bioelectronic device prepared by the method as described above,

wherein the surface of the nanomaterial is modified by the single-stranded nucleic acid handle, the single-stranded nucleic acid counter handle is introduced into the bioelectronic device, and when an electric field is applied to the first electrode and the second electrode of the bioelectronic device, the single-stranded nucleic acid counter handle is controllably exposed, and the nanomaterial modified by the single-stranded nucleic acid handle is controllably bound to the bioelectronic device.

According to yet another aspect of the disclosure, a synthesis circuit includes: the bioelectronic device as described above or the bioelectronic device prepared by the method as described above,

wherein nucleic acid sequences binding to different disease markers are provided to the bioelectronic device, the binding sequences are selectively converted when an electric field is applied to the first and second electrodes of the bioelectronic device, the converted binding sequences bind to the disease markers in the synthetic circuit, and a biomolecular output signal is generated to trigger the synthetic circuit, thereby obtaining a diagnostic result of the disease.

Drawings

The accompanying drawings, which are included to provide a further understanding of the disclosure and are incorporated in and constitute a part of this specification, illustrate exemplary embodiments of the disclosure and together with the description serve to explain the principles of the disclosure.

Fig. 1 shows a schematic view of a bioelectronic device according to an embodiment of the present disclosure.

Fig. 2 shows a flow diagram of a method of fabricating a bioelectronic device according to an embodiment of the present disclosure.

Fig. 3 shows a flow diagram of a method for self-assembled nucleic acid superstructure deposition according to one embodiment of the present disclosure.

FIG. 4 illustrates a flow chart of an electrode machining method according to one embodiment of the present disclosure.

Detailed Description

The present disclosure will be described in further detail with reference to the drawings and embodiments. It is to be understood that the specific embodiments described herein are for purposes of illustration only and are not to be construed as limitations of the present disclosure. It should be further noted that, for the convenience of description, only the portions relevant to the present disclosure are shown in the drawings.

It should be noted that the embodiments and features of the embodiments in the present disclosure may be combined with each other without conflict. Technical solutions of the present disclosure will be described in detail below with reference to the accompanying drawings in conjunction with embodiments.

Unless otherwise indicated, the illustrated exemplary embodiments/examples are to be understood as providing exemplary features of various details of some ways in which the technical concepts of the present disclosure may be practiced. Accordingly, unless otherwise indicated, features of the various embodiments may be additionally combined, separated, interchanged, and/or rearranged without departing from the technical concept of the present disclosure.

The use of cross-hatching and/or shading in the drawings is generally used to clarify the boundaries between adjacent components. As such, unless otherwise noted, the presence or absence of cross-hatching or shading does not convey or indicate any preference or requirement for a particular material, material property, size, proportion, commonality between the illustrated components and/or any other characteristic, attribute, property, etc., of a component. Further, in the drawings, the size and relative sizes of components may be exaggerated for clarity and/or descriptive purposes. While example embodiments may be practiced differently, the specific process sequence may be performed in a different order than that described. For example, two processes described consecutively may be performed substantially simultaneously or in reverse order to that described. In addition, like reference numerals denote like parts.

When an element is referred to as being "on" or "on," "connected to" or "coupled to" another element, it can be directly on, connected or coupled to the other element or intervening elements may be present. However, when an element is referred to as being "directly on," "directly connected to" or "directly coupled to" another element, there are no intervening elements present. For purposes of this disclosure, the term "connected" may refer to physically, electrically, etc., and may or may not have intermediate components.

The following description provides specific details such as material types, material thicknesses, and processing conditions in order to provide a thorough description of embodiments of the invention. However, it will be apparent to one skilled in the art that embodiments of the invention may be practiced without such specific details. Indeed, embodiments of the invention may be practiced in conjunction with conventional manufacturing techniques employed in the industry.

According to one embodiment of the present disclosure, a bioelectronic device is provided.

As shown in fig. 1, the bioelectronic device 100 may include a substrate 110, a first electrode 120, a second electrode 130, and a self-assembled nucleic acid superstructure 140.

The base material of the substrate 110 may include, but is not limited to, silicon dioxide, aluminum oxide, sapphire, germanium, gallium arsenide, alloys of silicon and germanium, or indium phosphide. In some cases, the substrate may include silicon nitride, carbon, and/or a polymer.

The base material of the substrate 110 may be inorganic (e.g., not containing carbon) or organic (e.g., containing carbon). In some cases, the substrate may comprise graphene and/or graphite.

In some embodiments, the substrate of the substrate 110 is a hybrid (e.g., comprises a mixture) of any two or more provided herein (e.g., a hybrid of an inorganic material and an organic material, or a hybrid of two or more different inorganic materials or organic materials). For example, the substrate may comprise a mixture of inorganic and organic materials, a mixture of two or more different inorganic materials, or a mixture of two or more different organic materials.

In some embodiments, the base material of the substrate 110 comprises a semiconductor material or a mixture of semiconductor materials. Semiconductor materials include, but are not limited to, group IV element semiconductors, group IV compound semiconductors, group VI element semiconductors, group III-V semiconductors, group II-VI semiconductors, group I-VII semiconductors, group IV-VI semiconductors, group V-VI semiconductors, group II-V semiconductors, oxides, layered semiconductors, magnetic semiconductors, organic semiconductors, charge transfer composites, and combinations thereof.

In some embodiments, the base material of substrate 110 comprises a group IV semiconductor material. Examples of group IV semiconductor materials for use in accordance with the present disclosure include, but are not limited to, diamond, silicon, germanium, gray tin, silicon carbide, and combinations thereof.

In some embodiments, the base material of substrate 110 comprises a group VI semiconductor material. Examples of group VI semiconductor materials for use in accordance with the present disclosure include, but are not limited to, sulfur, selenium ash, tellurium, and combinations thereof.

In some embodiments, the base material of substrate 110 comprises a III-V semiconductor material. Examples of III-V semiconductor materials for use in accordance with the present disclosure include, but are not limited to, cubic boron nitride, hexagonal boron nitride, boron phosphide, boron arsenide, aluminum nitride, aluminum phosphide, aluminum arsenide, aluminum antimonide, gallium nitride, gallium phosphide, gallium arsenide, gallium antimonide, indium nitride, indium phosphide, indium arsenide, indium antimonide, and combinations thereof.

In some embodiments, the base material of substrate 110 comprises a II-VI semiconductor material. Examples of group II-VI semiconductor materials for use in accordance with the present disclosure include, but are not limited to, cadmium selenide, cadmium sulfide, cadmium telluride, zinc oxide, zinc selenide, zinc sulfide, zinc telluride, cuprous chloride, copper sulfide, lead selenide, lead sulfide (II) lead telluride, tin sulfide, tin telluride, lead tin telluride, thallium germanium telluride, bismuth telluride, and combinations thereof.

In some embodiments, the base material of the substrate 110 comprises a group I-VII semiconductor material. Examples of group I-VII semiconductor materials for use in accordance with the present disclosure include, but are not limited to, cuprous chloride, cupric sulfide, and combinations of cuprous chloride and cupric sulfide.

In some embodiments, the base material of substrate 110 comprises a group IV-VI semiconductor material. Examples of group IV-VI semiconductor materials for use in accordance with the present disclosure include, but are not limited to, lead selenide, lead sulfide, lead telluride, tin sulfide, tin telluride, lead tin telluride, thallium germanium telluride, and combinations thereof.

In some embodiments, the base material of substrate 110 comprises a group V-VI semiconductor material. Examples of group IV-VI semiconductor materials for use in accordance with the present disclosure include, but are not limited to, bismuth telluride.

In some embodiments, the base material of substrate 110 comprises a II-V semiconductor material. Examples of group II-V semiconductor materials for use in accordance with the present disclosure include, but are not limited to, cadmium phosphide, cadmium arsenide, cadmium antimonide, zinc phosphide, zinc arsenide, zinc antimonide, and combinations thereof.

In some embodiments, the base material of the substrate 110 comprises an oxide. Examples of oxides for use in accordance with the present disclosure include, but are not limited to, anatase titanium dioxide, rutile titanium dioxide, brookite titanium dioxide, copper oxide, uranium dioxide, uranium trioxide, bismuth trioxide, tin dioxide, barium titanate, titanium acid, lithium niobate, lanthanum copper oxide, and combinations thereof.

In some embodiments, the base material of substrate 110 comprises a layered semiconductor. Examples of layered semiconductors for use in accordance with the present disclosure include, but are not limited to, lead iodide, molybdenum disulfide, gallium selenide, tin sulfide, bismuth sulfide, and combinations thereof.

In some embodiments, the base material of the substrate 110 comprises a magnetic semiconductor. Examples of magnetic semiconductors used in accordance with the present disclosure include, but are not limited to, gallium manganese arsenide, indium manganese arsenide, cadmium manganese telluride, lead manganese telluride, lanthanum calcium manganate, iron oxide, nickel oxide, europium sulfide, chromium bromide, and combinations thereof.

Other examples of semiconductor materials that may be used in accordance with the present disclosure include, but are not limited to, copper indium selenide, silver gallium sulfide, zinc silicon phosphide, arsenic sulfide, platinum silicide, iodine iodide, mercury iodide, thallium bromide, silver sulfide, iron disulfide, copper zinc tin sulfide, copper zinc antimony sulfide, and combinations thereof.

In some embodiments, the base material of substrate 110 comprises a chalcogenide. Chalcogenides are chemical compounds comprising at least one chalcogen anion and at least one more electropositive element. In some embodiments, the chalcogenide is a sulfide, selenide, or telluride.

In some embodiments, the substrate of substrate 110 comprises a film, such as a photoresist film, a chemical vapor deposition film, a semiconductor film, graphene, and/or other monolayer atomic film. In some embodiments, the substrate comprises a physical vapor deposition film, an atomic layer deposition film, and/or an ion implantation film.

In some embodiments, the substrate of substrate 110 is a polished silicon wafer, such as a silicon wafer treated with a plasma treatment, or a piranha solution (piranha solution).

The self-assembled nucleic acid superstructure 130 may be located above the substrate 110 and between the first electrode 120 and the second electrode 130. Therein, the self-assembled nucleic acid superstructure 140 may be deposited onto the substrate 110.

The first electrode 120 may be located on the substrate 110 and may be disposed on the substrate 110 in a manner that deposits to the surface of the self-assembled nucleic acid superstructure 140.

The second electrode 130 may be located on the substrate 110 and may be disposed on the substrate 110 in a manner that deposits to the surface of the self-assembled nucleic acid superstructure 140.

Wherein the self-assembled nucleic acid superstructure 140 is constructed from a specific amount, a specific sequence and a specific orientation of single-stranded nucleic acids, and controlled conversion of double-stranded nucleic acids to single-stranded nucleic acids in the specific amount, specific sequence and specific orientation is achieved by modulation of the electric field applied between the first electrode 120 and said second electrode 130. Where a particular amount refers to a predetermined amount of single-stranded nucleic acid and double-stranded nucleic acid, and a particular orientation refers to a controlled transformation in a certain direction or directions.

In an alternative embodiment, different voltages are applied via the first electrode 120 and the second electrode 130 to achieve a controlled conversion of double stranded nucleic acids to single stranded nucleic acids in a specific amount, a specific sequence and a specific orientation. Wherein the first electrode and the second electrode may be a source electrode and a drain electrode, respectively.

For example, in the design of the present disclosure, the current created based on the directional movement of ions inside the nucleic acid superstructure may not be used with the conductivity of the nucleic acid molecule itself. The voltage applied by the first electrode 120 and the second electrode 130 can be changed, so that ions in the nucleic acid superstructure are directionally moved under the action of an external electric field, the local ion concentration in the nucleic acid superstructure is reduced, the double-stranded nucleic acid can be dissociated, and the controllable conversion of the double-stranded nucleic acid and the single-stranded nucleic acid can be realized. The conformation transition (such as the switch state change of DNAAptamer switch) of biomolecules such as DNA/RNA/LNA/protein and the like under different electric field and ion movement conditions can also be utilized.

Nucleic acids of the present disclosure may include DNA, LNA, PNA, and RNA.

The basic principle of designing a self-assembling nucleic acid superstructure is to encode sequence complementarity in the nucleic acid strands such that by pairing the complementary fragments, the nucleic acid strands self-organize under appropriate physical conditions into a predefined nucleic acid superstructure.

In some embodiments, the nucleic acid superstructure is assembled from single-stranded nucleic acids, double-stranded nucleic acids, or a combination of single-stranded and double-stranded nucleic acids.

In some embodiments, the nucleic acid superstructure is assembled from a plurality of different nucleic acids (e.g., single-stranded nucleic acids). For example, a nucleic acid superstructure may be assembled from at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 nucleic acids. In some embodiments, the nucleic acid superstructure is assembled from at least 100, at least 200, at least 300, at least 400, at least 500, or more nucleic acids. The term "nucleic acid" encompasses "oligonucleotides," which are short single-stranded nucleic acids (e.g., DNA) of 10 nucleotides to 100 nucleotides in length. In some embodiments, the oligonucleotide is 10 to 20 nucleotides, 10 to 30 nucleotides, 10 to 40 nucleotides, 10 to 50 nucleotides, 10 to 60 nucleotides, 10 to 70 nucleotides, 10 to 80 nucleotides, or 10 to 90 nucleotides in length. In some embodiments, the oligonucleotide is 20 to 50, 20 to 75, or 20 to 100 nucleotides in length. In some embodiments, the oligonucleotide is 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleotides in length. In the case of a DNA origami structure, the longest nucleotide chain may have 8000 nucleotides.

The material of the first electrode 120 and the second electrode 130 is a metal material that can be used for deposition, such as titanium, gold, palladium, and the like. In addition, the first electrode 120 and the second electrode 130 may be deposited metal films or stacked metal films (composed of two or more metal films).

According to another embodiment of the present disclosure, the present disclosure also provides a method of manufacturing a bioelectronic device.

As shown in fig. 2, the preparation method may include step S10: synthesizing a self-assembled nucleic acid superstructure; step S20: depositing the self-assembled nucleic acid superstructure on a substrate; step S30: processing a first electrode and a second electrode on the surface of the deposited self-assembled nucleic acid superstructure.

Wherein the controllable conversion of double-stranded nucleic acid to single-stranded nucleic acid in a specific amount, a specific sequence and a specific orientation can be achieved by the regulation of the electric field applied between the first electrode and the second electrode.

In step S10, a self-assembled nucleic acid superstructure constructed from a specific number, a specific sequence and a specific orientation of single-stranded nucleic acids is designed to be synthesized. Wherein the synthesis can be carried out in an aqueous system.

Nucleic acids of the present disclosure include DNA, e.g., D-type DNA and L-type DNA, and/or RNA.

In some embodiments, the nucleic acids are combined in a single container (such as, but not limited to, a tube, well, or vial). The molar amount of nucleic acid used may depend on the frequency of each nucleic acid in the desired nucleic acid superstructure and the amount of superstructure desired.

In some embodiments, the DNA tile structure (DNA Brick) is placed in solution. The solution may be buffered by a buffer. The solution may further comprise metal cations such as Li, Mg, Na, K, Cs, Ca, Fe, Ni, and the like. The solution may also contain EDTA or other nuclease inhibitors to prevent nucleic acid degradation. Then incubation was performed. Wherein the synthesized nucleic acid superstructure can be used directly without purification. In addition to the DNA tile structure described above, other nucleic acid systems may be employed, such as DNA origami or RNA structures, etc.

Wherein the incubation can be performed under a stepwise temperature reaction condition. The incubation may be performed by selecting at least two temperatures in a temperature range of 100 to 20 ℃ and maintaining the temperatures at the at least two temperatures for a predetermined time, respectively.

The basic principle of designing a self-assembling nucleic acid superstructure is to encode sequence complementarity in the nucleic acid strands such that by pairing the complementary fragments, the nucleic acid strands self-organize under appropriate physical conditions into a predefined nucleic acid superstructure.

In some embodiments, the nucleic acid superstructure is assembled from single-stranded nucleic acids, double-stranded nucleic acids, or a combination of single-stranded and double-stranded nucleic acids.

In some embodiments, the nucleic acid superstructure is assembled from a plurality of different nucleic acids (e.g., single-stranded nucleic acids). For example, a nucleic acid superstructure can be assembled from at least 4, at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 nucleic acids. In some embodiments, the nucleic acid superstructure is assembled from at least 100, at least 200, at least 300, at least 400, at least 500, or more nucleic acids. The term "nucleic acid" encompasses "oligonucleotides," which are short single-stranded nucleic acids (e.g., DNA) of 10 nucleotides to 100 nucleotides in length. In some embodiments, the oligonucleotide is 10 to 20 nucleotides, 10 to 30 nucleotides, 10 to 40 nucleotides, 10 to 50 nucleotides, 10 to 60 nucleotides, 10 to 70 nucleotides, 10 to 80 nucleotides, or 10 to 90 nucleotides in length. In some embodiments, the oligonucleotide is 20 to 50, 20 to 75, or 20 to 100 nucleotides in length. In some embodiments, the oligonucleotide is 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleotides in length, and if in a DNA origami structure, the longest nucleotide strand can be 8000 nucleotides.

In step S20, the self-assembled nucleic acid superstructure is deposited on the substrate surface.

In this step S20, the following steps may be included, for example, see fig. 3, step S21: depositing the self-assembled nucleic acid superstructure on the substrate surface, step S22: incubating at room temperature for a period of time, step S23: removing the remaining solution (e.g., by wiping and/or by forced air flow), step S24: performing a desalting process for removing residual inorganic salts from the substrate, step S25: drying step, step S26: and imaging the self-assembled nucleic acid superstructure on the substrate.

Substrates used in accordance with the present disclosure may include, but are not limited to, silicon dioxide, aluminum oxide, sapphire, germanium, gallium arsenide, alloys of silicon and germanium, or indium phosphide. In some cases, the substrate may include silicon nitride, carbon, and/or a polymer.

The substrate may be inorganic (e.g., not containing carbon) or organic (e.g., containing carbon). In some cases, the substrate may comprise graphene and/or graphite.

In some embodiments, the substrate is a hybrid (e.g., comprising a mixture) of any two or more provided herein (e.g., a hybrid of an inorganic material and an organic material, or a hybrid of two or more different inorganic materials or organic materials). For example, the substrate may comprise a mixture of inorganic and organic materials, a mixture of two or more different inorganic materials, or a mixture of two or more different organic materials.

In some embodiments, the substrate comprises a semiconductor material or a mixture of semiconductor materials. Semiconductor materials include, but are not limited to, group IV element semiconductors, group IV compound semiconductors, group VI element semiconductors, group III-V semiconductors, group II-VI semiconductors, group I-VII semiconductors, group IV-VI semiconductors, group V-VI semiconductors, group II-V semiconductors, oxides, layered semiconductors, magnetic semiconductors, organic semiconductors, charge transfer composites, and combinations thereof.

In some embodiments, the substrate comprises a group IV semiconductor material. Examples of group IV semiconductor materials for use in accordance with the present disclosure include, but are not limited to, diamond, silicon, germanium, gray tin, silicon carbide, and combinations thereof.

In some embodiments, the substrate comprises a group VI semiconductor material. Examples of group VI semiconductor materials for use in accordance with the present disclosure include, but are not limited to, sulfur, selenium ash, tellurium, and combinations thereof.

In some embodiments, the substrate comprises a group III-V semiconductor material. Examples of III-V semiconductor materials for use in accordance with the present disclosure include, but are not limited to, cubic boron nitride, hexagonal boron nitride, boron phosphide, boron arsenide, aluminum nitride, aluminum phosphide, aluminum arsenide, aluminum antimonide, gallium nitride, gallium phosphide, gallium arsenide, gallium antimonide, indium nitride, indium phosphide, indium arsenide, indium antimonide, and combinations thereof.

In some embodiments, the substrate comprises a II-VI semiconductor material. Examples of group II-VI semiconductor materials for use in accordance with the present disclosure include, but are not limited to, cadmium selenide, cadmium sulfide, cadmium telluride, zinc oxide, zinc selenide, zinc sulfide, zinc telluride, cuprous chloride, copper sulfide, lead selenide, lead sulfide (II) lead telluride, tin sulfide, tin telluride, lead tin telluride, thallium germanium telluride, bismuth telluride, and combinations thereof.

In some embodiments, the substrate comprises a group I-VII semiconductor material. Examples of group I-VII semiconductor materials for use in accordance with the present disclosure include, but are not limited to, cuprous chloride, cupric sulfide, and combinations of cuprous chloride and cupric sulfide.

In some embodiments, the substrate comprises a group IV-VI semiconductor material. Examples of group IV-VI semiconductor materials for use in accordance with the present disclosure include, but are not limited to, lead selenide, lead sulfide, lead telluride, tin sulfide, tin telluride, lead tin telluride, thallium germanium telluride, and combinations thereof.

In some embodiments, the substrate comprises a group V-VI semiconductor material. Examples of group IV-VI semiconductor materials for use in accordance with the present disclosure include, but are not limited to, bismuth telluride.

In some embodiments, the substrate comprises a group II-V semiconductor material. Examples of group II-V semiconductor materials for use in accordance with the present disclosure include, but are not limited to, cadmium phosphide, cadmium arsenide, cadmium antimonide, zinc phosphide, zinc arsenide, zinc antimonide, and combinations thereof.

In some embodiments, the substrate comprises an oxide. Examples of oxides for use in accordance with the present disclosure include, but are not limited to, anatase titanium dioxide, rutile titanium dioxide, brookite titanium dioxide, copper oxide, uranium dioxide, uranium trioxide, bismuth trioxide, tin dioxide, barium titanate, titanium acid, lithium niobate, lanthanum copper oxide, and combinations thereof.

In some embodiments, the substrate comprises a layered semiconductor. Examples of layered semiconductors for use in accordance with the present disclosure include, but are not limited to, lead iodide, molybdenum disulfide, gallium selenide, tin sulfide, bismuth sulfide, and combinations thereof.

In some embodiments, the substrate comprises a magnetic semiconductor. Examples of magnetic semiconductors used in accordance with the present disclosure include, but are not limited to, gallium manganese arsenide, indium manganese arsenide, cadmium manganese telluride, lead manganese telluride, lanthanum calcium manganate, iron oxide, nickel oxide, europium sulfide, chromium bromide, and combinations thereof.

Other examples of semiconductor materials that may be used in accordance with the present disclosure include, but are not limited to, copper indium selenide, silver gallium sulfide, zinc silicon phosphide, arsenic sulfide, platinum silicide, iodine iodide, mercury iodide, thallium bromide, silver sulfide, iron disulfide, copper zinc tin sulfide, copper zinc antimony sulfide, and combinations thereof.

In some embodiments, the substrate comprises a chalcogenide. Chalcogenides are chemical compounds comprising at least one chalcogen anion and at least one more electropositive element. In some embodiments, the chalcogenide is a sulfide, selenide, or telluride.

In some embodiments, the substrate comprises a film, such as a photoresist film, a chemical vapor deposited film, a semiconductor film, graphene, and/or other monolayer atomic film. In some embodiments, the substrate comprises a physical vapor deposition film, an atomic layer deposition film, and/or an ion implantation film.

The substrate should include organic thin films and polymers, such as PMMA, PDMS, etc

In some embodiments, the substrate is a polished silicon wafer, such as a silicon wafer treated with a plasma treatment, or a piranha solution (piranha solution).

In the present disclosure, the nucleic acid superstructure may be deposited at a fixed location, and may also be deposited with reference to an alignment mark.

In determining the location of the nucleic acid superstructure relative to the alignment marks, the alignment marks may be processed prior to deposition of the nucleic acid superstructure.

In the process of processing the alignment mark, firstly, a photoresist layer is coated on a substrate in a rotating mode, then a fine alignment mark pattern is written, and the alignment mark pattern is developed through a developing solution. Depositing a metal film or stacking metal films, and then stripping the photoresist layer using a stripping solution, where the stripping can be performed without using an ultrasonic means. And then cleaning and drying. The cleaning solution may be diluted methanol/ethanol/isopropanol (30-95 vol%).

In step S30, a first electrode and a second electrode are machined at specific locations on the surface of the self-assembled nucleic acid superstructure.

See, for example, fig. 4, where step S30 may include: step S31: spin coating a photoresist layer to the surface of the self-assembled nucleic acid superstructure, step S32: writing the pattern of the first electrode and the second electrode, step S33: developing the first electrode and the second electrode pattern by the developing solution, step S34: depositing a metal film or stacking metal films (e.g., two or more stacked metal films, wherein the metal material may be a metal material that can be used for deposition), step S34: the photoresist layer is stripped using a stripping solution, where the stripping can be performed without using an ultrasonic means. And then cleaning and drying. The cleaning solution may be diluted methanol/ethanol/isopropanol (30-95 vol%).

According to still another embodiment of the present disclosure, there is also provided a method for controlled transformation of double-stranded nucleic acid and single-stranded nucleic acid by the above-described bioelectronic device or the bioelectronic device prepared by the above-described method, wherein the controlled transformation of double-stranded nucleic acid and single-stranded nucleic acid in a specific amount, a specific sequence and a specific orientation is achieved by electric field regulation between the first electrode and the second electrode. Wherein the first electrode and the second electrode may be a source electrode and a drain electrode, respectively.

For example, in the design of the present disclosure, the current created based on the directional movement of ions inside the nucleic acid superstructure may not be used with the conductivity of the nucleic acid molecule itself. The voltage applied by the first electrode and the second electrode can be changed, so that ions in the nucleic acid superstructure directionally move under the action of an external electric field, the local ion concentration in the nucleic acid superstructure is reduced, the double-stranded nucleic acid can be dissociated, and the controllable conversion of the double-stranded nucleic acid and the single-stranded nucleic acid is realized.

Controlled conversion in the present disclosure, dissociation of different double-stranded nucleic acids can be achieved when the applied voltage is different. For example, when a lower voltage is applied, dissociation of a double-stranded nucleic acid with weaker binding can be achieved, so that different double-stranded nucleic acids can be dissociated according to different voltages. Because the self-assembly nucleic acid superstructure has intrinsic quantitative, sequence-fixed and space-fixed orientation characteristics, the controllable conversion from double-stranded nucleic acid of the self-assembly nucleic acid superstructure to single-stranded nucleic acid can be realized.

Therefore, the present disclosure provides a novel electrically controlled nucleic acid switch, which can use an electric field to regulate and control the controllable expression of single-stranded nucleic acid, and realize the controllable transformation of the quantification, the fixed sequence and the fixed spatial orientation of double-stranded nucleic acid and single-stranded nucleic acid.

In the present disclosure, a chemical modification of an electronic material is not used to achieve quantitative, sequence-fixed, and orientation-fixed controllable transformation (for example, chemical modification of single-stranded nucleic acid at a specific position on the surface of a carbon nanotube), so as to avoid the influence of random uncontrollable property of chemical modification on the quantity and spatial orientation of the single-stranded nucleic acid, which may be uncontrollable distributed on the surface of the electronic material due to the random distribution property of the modification chemical reaction itself, and thus the quantity and relative spatial position of the single-stranded nucleic acid may not be controllable.

According to a further embodiment of the present disclosure, there is also provided a system for selectively controlling protein binding, in which the above bioelectronic device or the bioelectronic device prepared by the above method is included.

In the bioelectronic devices (electronically controlled nucleic acid switches) described above, controlled conversion of a specific amount, a specific sequence and a specific orientation of double-stranded nucleic acids and single-stranded nucleic acids can be performed by electric field regulation by two electrodes. Thus, protein binding can be selectively controlled by a system for controlling protein binding including the bioelectronic device. For example, in the case of DNA, the DNA single-stranded sequence may be designed to have a specific aptamer or to contain a protein binding site. These single-stranded DNAs bound to proteins remain in a double-stranded form during DNA formation.

When an electric field is applied to both electrodes of the bioelectronic device, the aptamer sequence or protein binding sequence is exposed and selectively captures the protein only at the designed site in solution. Moreover, the combination of multiple heterogeneous proteins can be demonstrated in the same switch by introducing different sequences of protein-specific single-stranded DNA. In this way, the nucleic acid switch may sense the presence of the protein.

There is also provided, in accordance with yet another embodiment of the present disclosure, a system for selectively controlling nanomaterial incorporation, in which the bioelectronic device described above or a bioelectronic device prepared by the above method is included.

Similar to the above-described protein binding, the above-described bioelectronic devices (electronically controlled nucleic acid switches) can be used to capture nanomaterials, such as gold nanoparticles and carbon nanotubes, to control binding of the nanomaterials. The surface of the nanomaterial may be modified using, for example, a single-stranded DNA handle (DNA handle). A single-stranded DNA anti-handle (complementary sequence of the DNA handle) is introduced (building a nucleic acid superstructure) into the switch, and when an electric field is applied across the two electrodes of the bioelectronic device, the single-stranded DNA anti-handle is exposed and the nanomaterial will selectively bind to the switch.

According to still another embodiment of the present disclosure, there is also provided a synthesis circuit including the above bioelectronic device or the bioelectronic device prepared by the above method.

Synthetic circuits formed by DNA or RNA can be used for disease diagnosis (e.g., single nucleotide polymorphisms). Wherein different disease markers can be selectively identified by simply changing the binding sequence of the synthesis circuit while keeping the decision unit unchanged in the synthesis loop.

To use the bioelectronic devices (nucleic acid switches) of the present disclosure in a synthetic circuit, nucleic acid sequences that bind to different disease markers are immobilized into the nucleic acid switches. When an electrical signal (current or voltage) is applied to the two electrodes of the nucleic acid switch, different binding sequences can be selectively activated. The exposed binding sequences (which are selectively activated) bind to disease markers in the synthetic circuit, generating a biomolecule output signal (e.g., single-stranded DNA or protein) to trigger the synthetic circuit and report a diagnostic result. Finally, because the turning on or off of multiple binding sequences can be selectively controlled, multiple binding sequences can be activated simultaneously, thereby triggering the response of the synthesis circuit to multiple inputs and improving diagnostic accuracy.

In the description herein, reference to the description of the terms "one embodiment/mode," "some embodiments/modes," "example," "specific example," or "some examples," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment/mode or example is included in at least one embodiment/mode or example of the application. In this specification, the schematic representations of the terms used above are not necessarily intended to be the same embodiment/mode or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments/modes or examples. Furthermore, the various embodiments/aspects or examples and features of the various embodiments/aspects or examples described in this specification can be combined and combined by one skilled in the art without conflicting therewith.

Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In the description of the present application, "plurality" means at least two, e.g., two, three, etc., unless specifically limited otherwise.

It will be understood by those skilled in the art that the foregoing embodiments are merely for clarity of illustration of the disclosure and are not intended to limit the scope of the disclosure. Other variations or modifications may occur to those skilled in the art, based on the foregoing disclosure, and are still within the scope of the present disclosure.