Binary digital encryption operation method based on DNA nano switch

Abstract

The invention discloses a binary digital encryption operation method based on a DNA nano switch. The invention belongs to the technical field of information storage and encryption of biomolecule self-assembly materials, and is characterized in that four basic numbers 1, 2, 4 and 8 are prepared through different annular DNA nano switches on the basis of a strategy of'protection first and deprotection later '; secondly, mixing basic numbers according to the operation required to be carried out, and triggering operation encryption by using RNase A ribonucleic acid endonuclease; and finally, adding a connecting chain T to trigger operation, and reading an operation result through an agarose gel electrophoresis technology. A simple and low-cost information storage and digital operation scheme is provided, the DNA nano switch combines response characteristics of biomolecules with information encryption requirements, a scheme is provided for high-safety, high-density and biocompatible data protection, and a possible new path is provided for a low-energy-consumption and high-secrecy biological computer and precise medical operation; meanwhile, the DNA nano switch has rewritability, and the method has very high expansibility and can be possibly applied to more complex information storage and logic operation scenes in the future.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomolecular self-assembled material information storage and encryption, and specifically refers to a binary digital encryption operation method based on a DNA nanoswitch. Background Art

[0002] A DNA nanoswitch is a type of DNA nanostructure constructed based on the principle of DNA origami: using a single-stranded M13mp18 DNA as the backbone, it self-assembles with complementary short oligonucleotides to form a simple DNA linear double-stranded structure. A certain number of short oligonucleotides are designed according to experimental needs, with one end connected and paired with the backbone, and the other end being an unpaired and exposed sequence, and this part of the sequence can be complementary paired with the connecting strand. When the detection strand is complementary paired with the target strand, the DNA nanoswitch undergoes a topological deformation from a linear structure to a circular structure. Circular DNA nanoswitches have different migration rates in agarose gel electrophoresis due to different configurations and will show bands at different positions in the gel electrophoresis. Therefore, DNA circular nanoswitches with different configurations can be used as information carriers to carry the encoding and transmission of different information.

[0003] K. Halvorsen et al. designed and synthesized circular DNA nanoswitches with different configurations, and successfully achieved the encoding and storage of different letter information by using their different migration rates in agarose gel electrophoresis. Then, using RNase H ribonuclease for specific enzymatic cleavage to digest the RNA in the DNA-RNA hybrid region, the circular DNA nanoswitch was transformed into a linear structure, thereby performing "encryption" processing on the input information; finally, a single-stranded DNA was added to "decrypt" and read the transmitted information. Based on this strategy, the storage, encryption, and transmission of information were successfully realized. The scientific research team of Shaanxi Visono borrowed the strategy of "protection first - deprotection later" in organic chemistry, and proposed a strategy of first using a short RNA protection strand to prevent the DNA nanoswitch from forming a ring, and then performing information encryption and decryption through specific enzymatic cleavage methods.

[0004] According to whether the nanoswitch forms a ring, the linear structure and circular structure of the DNA nanoswitch are respectively defined as "0 and 1" in the binary counting system. Based on the strategy of "protection first - deprotection later", we successfully constructed the numbers 1, 2, 4, and 8 represented in binary. Using these 4 numbers as basic modules, and then relying on the specific enzymatic cleavage effect of RNaseA, through encryption and decryption for the expression, erasure, and rewriting of digital information, we realized the encryption operation of binary numbers within 15, providing new ideas and methods for the application of nanoswitches. Summary of the Invention

[0005] The objective of the present invention is to provide a binary digital encryption operation method based on DNA nanoswitches. The present invention is based on the "protection first - deprotection later" strategy, and uses different circular DNA nanoswitches to prepare the four basic numbers 1, 2, 4, and 8. Secondly, according to the operations to be performed, the basic numbers are mixed, and RNaseA endonuclease is used to trigger the operation encryption. Finally, a ligation strand T is added to trigger the operation and the operation result is read through agarose gel electrophoresis technology; a simple and low - cost information storage and digital operation solution is provided. By combining the response characteristics of biomolecules with the information encryption requirements, DNA nanoswitches provide a solution for highly secure, high - density, and biocompatible data protection, and provide a possible new path for low - energy - consumption, highly confidential biological computing and precision medical operations; at the same time, DNA nanoswitches have rewritability, and the present invention has strong scalability and may be applied to more complex information storage and logical operation scenarios in the future.

[0006] To achieve the above objective, the technical solution adopted by the present invention is as follows: A binary digital encryption operation method based on DNA nanoswitches, and the encryption operation strategy includes the following steps:

[0007] (1) Prepare linear DNA nanoswitches;

[0008] (2) According to the "protection first - deprotection later" strategy, prepare circular DNA nanoswitches corresponding to the basic numbers;

[0009] (3) Complete the storage of digital information by selectively mixing circular DNA nanoswitches;

[0010] (4) Use RNaseA nuclease to perform enzymatic digestion on the mixed solution to achieve the encryption of digital operations;

[0011] (5) After adding the target strand and incubating, read the operation result through agarose gel electrophoresis.

[0012] Further, the preparation process of the linear DNA nanoswitches in step (1) is specifically as follows: Use M13mp18 linear single - strand as the backbone strand to prepare linear single - strand DNA with a concentration of 250 ng / μL by enzymatic digestion; design and synthesize a mixture containing 149 staple strands and 5 detection strands D x ; Mix the staple strands in equal proportions to form a staple strand mixture M1, and mix the detection strands in equal proportions to form a detection strand mixture M2; Mix the staple strand - detection strand mixture M3 with the M13mp18 linear single - strand to form a mixture M4; Place the mixture M4 in a PCR instrument for an annealing program and then dilute it into 1×PBS buffer for standby.

[0013] Further, the preparation process of the circular DNA nanoswitch in step (2) is specifically as follows: Design the RNA ligation strand RNA 0-x Each strand contains 24 bases. 12 bases at one end are connected to the detection strand D0, and 12 bases at the other end are connected to the detection strand D x ; Through the RNA 0-x strand and the base complementary pairing of the detection strands D0 and D x to form circular DNA nanoswitches Loop 1, Loop 2, Loop 3, and Loop 4 with different configurations, corresponding to the decimal numbers 1, 2, 4, and 8 respectively.

[0014] Preferably, the design of the detection strand D x in step (2) follows the distance control principle. The pairing position of the detection strand D0 and the M13mp18 linear single strand is designed at the 2000th base from the 3' end of the M13mp18 linear single strand. The distance between the detection strand D1 and D0 is 600 bases, the distance between D2 and D0 is 1200 bases, the distance between D3 and D0 is 1800 bases, and the distance between D4 and D0 is 2400 bases.

[0015] Further, the storage of digital information in step (3) is achieved by selectively mixing the circular DNA nanoswitches corresponding to the basic digits; among them, the number 11 is represented by mixing the circular DNA nanoswitches corresponding to RNA 0-1 , RNA 0-2 , RNA 0-4 , and the number 15 is represented by mixing the circular DNA nanoswitches corresponding to RNA 0-1 , RNA 0-2 , RNA 0-3 , RNA 0-4 .

[0016] Further, the encryption process of digital operation in step (4) is completed by incubating at 37 °C for 10 minutes after adding RNaseA nuclease, specifically digesting the RNA 0-x strand to transform the circular DNA nanoswitch into a linear structure.

[0017] Further, the design of the RNA ligation strand RNA 0-x in step (4) is that each strand contains 24 bases. 12 bases at one end are connected to the detection strand D0, and 12 bases at the other end are connected to the detection strand D x to form a circular DNA nanoswitch.

[0018] Further, the reading of the operation result in step (5) is completed after incubating with the target strand T x ; The target strand T xis designed such that each strand contains 24 bases, with 12 bases at one end connected to the detection strand D0 and 12 bases at the other end connected to the detection strand D x connected.

[0019] Furthermore, in the step (5), the binary digital encryption operation strategy realizes the reading of the operation result by observing the position of the circular DNA nanoswitch on the gel diagram through agarose gel electrophoresis. The agarose gel electrophoresis conditions are set as 1.2% agarose concentration, 0.5×TBE buffer, 60V voltage, and 1.5 hours of running gel time.

[0020] Furthermore, in the step (5), the target strand T x is designed such that each strand contains 24 bases, with 12 bases at one end connected to the detection strand D0 and 12 bases at the other end connected to the detection strand D x connected, and is used to trigger the operation and decryption.

[0021] The beneficial effects obtained by the present invention with the above structure are as follows: (2) The present invention is based on the strategy of "protection first - deprotection later", and prepares four basic numbers 1, 2, 4, and 8 through different circular DNA nanoswitches; (2) The present invention mixes the basic numbers according to the operations required, uses RNaseA ribonuclease to trigger the operation encryption; finally, adds the connecting strand T to trigger the operation and reads the operation result through agarose gel electrophoresis technology; (3) The present invention provides a simple and low - cost information storage and digital operation solution. By combining the response characteristics of biomolecules with the information encryption requirements, the DNA nanoswitch provides a solution for high - security, high - density, and biocompatible data protection, and provides a possible new path for low - energy - consumption, high - confidentiality biological computing and precision medical operations; (4) The DNA nanoswitch in the present invention has rewritability, and the present invention has strong scalability and may be applied to more complex information storage and logical operation scenarios in the future. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 Example 1 (Addition operation);

[0023] Figure 2 Example 2 (Subtraction operation);

[0024] Figure 3 Example 3 (Multiplication operation);

[0025] Figure 4 Example 4 (Division operation).

[0026] The drawings are used to provide a further understanding of the present invention, and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation to the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0027] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments; based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0028] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those familiar to those skilled in the art. Furthermore, any methods and materials similar or equivalent to those described herein can be applied to the present invention. The preferred embodiments and materials described herein are for illustrative purposes only and are not intended to limit the scope of this application.

[0029] The experimental methods in the following examples are conventional methods unless otherwise specified; the experimental materials and reagents used in the following examples are purchased from commercial channels unless otherwise specified.

[0030] Example 1

[0031] Example DNA information addition based on nano-switches

[0032] (1) The binary codes corresponding to the decimal numbers "0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15" are: 0000, 0001, 0010, 0011, 0100, 0101, 0110, 0111, 1000, 1001, 1010, 1011, 1100, 1101, 1110, 1111;

[0033] (2) By constructing a 4-bit DNA nanoswitch, adding R 0-x Then four circular DNA nanoswitches Loop 4, Loop 3, Loop 2, and Loop 1 will be formed. After their corresponding circular and linear structures are mapped to binary 1 and 0 respectively, the circular DNA nanoswitches Loop 4, Loop 3, Loop 2, and Loop 1 from top to bottom correspond to the decimal numbers 8, 4, 2, and 1 respectively; the basic numbers 1, 2, 4, and 8 are mapped to the decimal numbers 8, 4, 2, and 1 respectively through RNA. 0-x Protective chain (RNA 0-1 , RNA 0-2 , RNA 0-3 , RNA 0-4 ) D0 and D x The connections form a ring-shaped nanoswitch representation;

[0034] (3) Taking the number 11 as an example, if the arithmetic formula for expressing the number 11 based on 4 basic numbers is 11 = 1 + 2 + 8; then the required RNA protection chain addition scheme is shown in Table 1;

[0035] Table 1 RNA protection chain addition scheme for the number 11

[0036]

[0037] (4) Aliquot a part of Samples 1, 2, and 3 in step (3) respectively, and incubate at 25 °C for 30 min; mix the remaining parts in a PCR tube, add RNase A, incubate at 37 °C for 10 min in a PCR instrument to digest RNA, and incubate at 25 °C for 30 min;

[0038] (5) Add the target strands T1, T2, and T4 to the mixed solution in step (4), mix well, and incubate at 25 °C for 30 min in a PCR instrument to complete the digital operation; read the operation result by agarose gel electrophoresis;

[0039] (6) Taking the number 15 as an example, if the arithmetic formula for expressing the number 15 based on 4 basic numbers is 15 = 1 + 2 + 4 + 8; then the required RNA protection chain addition scheme is shown in Table 2;

[0040] (7) Aliquot a part of Samples 1, 2, 3, and 4 in step (6) respectively, and incubate at 25 °C for 30 min; mix the remaining parts in a PCR tube, add RNase A, incubate at 37 °C for 10 min in a PCR instrument to digest RNA, and incubate at 25 °C for 30 min;

[0041] (8) Add the target strands T1, T2, T3, and T4 to the mixed solution in step (7) and mix well, incubate at 25 °C for 30 min in a PCR instrument to complete the digital operation; read the operation result by agarose gel electrophoresis;

[0042] Table 2 RNA protection chain addition scheme for the number 15

[0043]

[0044] (9) Verify by agarose gel electrophoresis. The electrophoresis buffer is 0.5×TBE, prepare an agarose gel with a concentration of 1.2%, select a Bio-Rad electrophoresis instrument and perform electrophoresis at 60 V for 1.5 h, and observe and read the operation result in a gel imaging system.

[0045] Example 2

[0046] Example DNA information performs subtraction operations based on nanoswitches

[0047] (1) The binary codes corresponding to the decimal digits "0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15" are: 0000, 0001, 0010, 0011, 0100, 0101, 0110, 0111, 1000, 1001, 1010, 1011, 1100, 1101, 1110, 1111;

[0048] (2) By constructing a 4 - bit DNA nanoswitch and adding R 0-x four circular DNA nanoswitches Loop 4, Loop 3, Loop 2, and Loop 1 will be formed. After corresponding their respective circular and linear structures to binary 1 and 0, the top - down circular DNA nanoswitches Loop 4, Loop 3, Loop 2, and Loop 1 correspond to decimal numbers 8, 4, 2, and 1 respectively; The basic numbers 1, 2, 4, and 8 are used by RNA 0-x protection strands (RNA 0-1 , RNA 0-2 , RNA 0-3 , RNA 0-4 ) to connect D0 and D x to form a circular nanoswitch representation;

[0049] (3) Taking the number 2 as an example, if the arithmetic formula for expressing the number 2 based on 4 basic numbers is 2 = 6 - 4; then the required RNA protection strand addition scheme is shown in Table 3;

[0050] Table 3 RNA protection strand addition scheme for number 2

[0051]

[0052] (4) Aliquot a part of Samples 1 and 2 in step (3) and incubate at 25 °C for 30 min; Mix the remaining parts in a single PCR tube, add RNase A, incubate in a PCR instrument at 37 °C for 10 min to digest RNA, and then incubate at 25 °C for 30 min;

[0053] (5) Add T2 in step (4), mix well and incubate in a PCR instrument at 25 °C for 30 min to complete the digital operation; Read the operation result through agarose gel electrophoresis;

[0054] (6) Taking the number 13 as an example, if the arithmetic formula for expressing the number 13 based on 4 basic numbers is 13 = 15 - 2; then the required RNA protection strand addition scheme is shown in Table 4;

[0055] Table 4 RNA protection strand addition scheme for number 13

[0056]

[0057] (7) Aliquot a part of Samples 1 and 2 from step (6) and incubate at 25 °C for 30 min; mix the remaining parts in one PCR tube, add RNase A, incubate at 37 °C for 10 min in a PCR instrument to digest RNA, and then incubate at 25 °C for 30 min;

[0058] (8) Add T1, T3, and T4 in step (7), incubate at 25 °C for 30 min in a PCR instrument to complete digital operation; read the operation result by agarose gel electrophoresis;

[0059] (9) Verify by agarose gel electrophoresis. The electrophoresis buffer is 0.5×TBE. Prepare an agarose gel with a concentration of 1.2%, use a Bio-Rad electrophoresis instrument at 60 V for electrophoresis for 1.5 h, and observe and read the operation result in a gel imaging system.

[0060] Example 3

[0061] The example DNA information performs multiplication operations based on a nanoswitch

[0062] (1) The binary codes corresponding to the decimal numbers "0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15" are: 0000, 0001, 0010, 0011, 0100, 0101, 0110, 0111, 1000, 1001, 1010, 1011, 1100, 1101, 1110, 1111;

[0063] (2) By constructing a 4-bit DNA nanoswitch and adding R 0-x Four circular DNA nanoswitches Loop 4, Loop 3, Loop 2, and Loop 1 will be formed. After corresponding their respective circular and linear structures to 1 and 0 in binary, the circular DNA nanoswitches Loop 4, Loop 3, Loop 2, and Loop 1 from top to bottom correspond to the decimal numbers 8, 4, 2, and 1 respectively; the basic numbers 1, 2, 4, and 8 are represented by connecting D0 and D 0-x through RNA 0-1 protection strands (RNA 0-2 、RNA 0-3 、RNA 0-4 ); x to form a circular nanoswitch representation;

[0064] (3) Taking the number 8 as an example, if the operation formula for expressing the number 8 based on 4 basic numbers is 8 = 1×2×4; the required RNA protection strand addition scheme is shown in Table 5;

[0065] Table 5 RNA protection strand addition scheme for the number 8

[0066]

[0067] (4) Aliquot a part of Samples 1, 2, and 3 from step (3) and incubate at 25 °C for 30 min; Mix the remaining parts in one PCR tube, add RNase A, incubate at 37 °C for 10 min in a PCR instrument to digest RNA, and then incubate at 25 °C for 30 min;

[0068] (5) Add the target strand T4 in step (4), mix well and incubate at 25 °C for 30 min in a PCR instrument; Complete the digital operation; Read the operation result by agarose gel electrophoresis;

[0069] (6) Taking the number 14 as an example again, if the operation formula for expressing the number 14 based on 4 basic numbers is 14 = 1 × 2 × 7; Then the required RNA protection strand addition scheme is shown in Table 6 below;

[0070] Table 6 RNA protection strand addition scheme for the number 14

[0071]

[0072] (7) Aliquot a part of Samples 1, 2, and 3 from step (6) and incubate at 25 °C for 30 min; Mix the remaining parts in one PCR tube, add RNase A, incubate at 37 °C for 10 min in a PCR instrument to digest RNA, and then incubate at 25 °C for 30 min;

[0073] (8) Add the target strands T2, T3, and T4 in step (7); Mix well and incubate at 25 °C for 30 min in a PCR instrument to complete the digital operation; Read the operation result by agarose gel electrophoresis;

[0074] (9) Verify by agarose gel electrophoresis. The electrophoresis buffer is 0.5×TBE. Prepare an agarose gel with a concentration of 1.2%. Use a Bio-Rad electrophoresis instrument at 60 V for electrophoresis for 1.5 h, and observe and read the operation result in a gel imaging system.

[0075] Example 4

[0076] Example DNA information performs division operations based on nanoswitches

[0077] (1) The binary codes corresponding to the decimal numbers "0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15" are: 0000, 0001, 0010, 0011, 0100, 0101, 0110, 0111, 1000, 1001, 1010, 1011, 1100, 1101, 1110, 111;

[0078] (2) By constructing a 4 - bit DNA nanoswitch and adding R 0-x Four circular DNA nanoswitches, Loop 4, Loop 3, Loop 2, and Loop 1, will be formed. After corresponding their respective circular and linear structures to binary 1 and 0 respectively, then the top - down circular DNA nanoswitches Loop 4, Loop 3, Loop 2, and Loop 1 correspond to decimal numbers 8, 4, 2, and 1 respectively; the basic numbers 1, 2, 4, and 8 are connected through RNA 0-x protection strands (RNA 0-1 、RNA 0-2 、RNA 0-3 、RNA 0-4 ) to form a representation of the circular nanoswitch by connecting D0 and D x ;

[0079] Table 7 Addition scheme of the protection strand for the number 4

[0080]

[0081]

[0082] (3) Taking the number 4 as an example, if the arithmetic formula for expressing the number 4 based on 4 basic numbers is 4 = 12÷3; then the required RNA protection strand addition scheme is shown in Table 7;

[0083] (4) Aliquot a part of samples 1 and 2 in step (3) and incubate at 25 °C for 30 min; mix the remaining parts in a single PCR tube, add RNase A, incubate at 37 °C for 10 min in a PCR instrument to digest RNA, and then incubate at 25 °C for 30 min;

[0084] (5) Add the target strand T3 in step (4); after mixing, incubate at 25 °C for 30 min in a PCR instrument to complete the digital operation; read the operation result by agarose gel electrophoresis;

[0085] (6) Then taking the number 5 as an example, if the arithmetic formula for expressing the number 5 based on 4 basic numbers is 5 = 10÷2; then the required RNA protection strand addition scheme is shown in Table 8

[0086] Table 8 Addition scheme of the RNA protection strand for the number 5

[0087]

[0088] (7) Aliquot a portion of Samples 1 and 2 from step (6) and incubate them at 25 °C for 30 min; mix the remaining portions in one PCR tube, add RNase A, incubate at 37 °C for 10 min in a PCR instrument to digest RNA, and then incubate at 25 °C for 30 min;

[0089] (8) Add target strands T1 and T3 in step (7), mix well, and incubate at 25 °C for 30 min in a PCR instrument to complete digital arithmetic; read the arithmetic result by agarose gel electrophoresis;

[0090] (9) Verify by agarose gel electrophoresis. The electrophoresis buffer is 0.5×TBE. Prepare an agarose gel with a concentration of 1.2%, and use a Bio-Rad electrophoresis instrument to perform electrophoresis at 60 V for 1.5 h. Observe and read the arithmetic result in a gel imaging system.

[0091] Although the embodiments of the present invention have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.

[0092] The above describes the present invention and its embodiments. Such a description is not restrictive. What is shown in the drawings is only one of the embodiments of the present invention, and the actual structure is not limited thereto. In general, if those of ordinary skill in the art are inspired by it and, without departing from the purpose of the present invention, design similar structural forms and embodiments to this technical solution without creative efforts, they should all fall within the protection scope of the present invention.

Claims

1. A binary digital encryption operation method based on a DNA nanoswitch, characterized in that: The encryption operation strategy includes the following steps: (1) Prepare linear DNA nanoswitches; (2) Prepare circular DNA nanoswitches corresponding to basic digits according to the "protection first - deprotection later" strategy; (3) Complete the storage of digital information by selectively mixing circular DNA nanoswitches; (4) Use RNaseA nuclease to perform enzymatic digestion on the mixed solution to achieve encryption of digital operations; (5) After adding the target strand and incubating, read the operation result through agarose gel electrophoresis.

2. The binary digital encryption operation method based on a DNA nanoswitch according to claim 1, wherein: The preparation process of the linear DNA nanoswitch in step (1) is specifically as follows: Using M13mp18 linear single-strand as the backbone strand, prepare linear single-stranded DNA with a concentration of 250 ng / μL by enzymatic digestion; design and synthesize a mixture containing 149 staple strands and 5 detection strands D x ; Mix the staple strands in equal proportions to form staple strand mixture M1, and mix the detection strands in equal proportions to form detection strand mixture M2; Mix the staple strand-detection strand mixture M3 with the M13mp18 linear single-strand to form mixture M4; Place mixture M4 in a PCR instrument for an annealing program and then dilute it in 1×PBS buffer for standby.

3. The binary digital encryption operation method based on a DNA nanoswitch according to claim 2, wherein: In the step (2), the preparation process of the circular DNA nanoswitch is specifically as follows: Design an RNA linker RNA 0-x Each strand contains 24 bases. 12 bases at one end are connected to the detection strand D0, and 12 bases at the other end are connected to the detection strand D x ; Through the base complementary pairing of the RNA 0-x strand with the detection strands D0 and D x , circular DNA nanoswitches Loop 1, Loop 2, Loop 3, and Loop 4 with different configurations are formed, corresponding to the decimal numbers 1, 2, 4, and 8 respectively.

4. A binary digital encryption operation method based on a DNA nanoswitch according to claim 3, characterized in that: The detection strand D in the step (2) x is designed according to the distance control principle. The pairing position of the detection strand D0 and the M13mp18 linear single strand is designed at the 2000th base from the 3' end of the M13mp18 linear single strand. The distance between the detection strand D1 and D0 is 600 bases, the distance between D2 and D0 is 1200 bases, the distance between D3 and D0 is 1800 bases, and the distance between D4 and D0 is 2400 bases.

5. A binary digital encryption operation method based on a DNA nanoswitch according to claim 4, characterized in that: The storage of the digital information in step (3) is achieved by selectively mixing circular DNA nanoswitches corresponding to basic digits; among them, the number 11 is represented by mixing circular DNA nanoswitches corresponding to RNA 0-1 , RNA 0-2 , RNA 0-4 , and the number 15 is represented by mixing circular DNA nanoswitches corresponding to RNA 0-1 , RNA 0-2 , RNA 0-3 , RNA 0-4 .

6. The binary digital encryption operation method based on a DNA nanoswitch according to claim 5, characterized in that: The encryption process of the digital operation in step (4) is completed by incubating with RNase A nuclease at 37 °C for 10 minutes, specifically digesting the RNA 0-x strand to transform the circular DNA nanoswitch into a linear structure.

7. A binary digital encryption operation method based on a DNA nanoswitch according to claim 6, characterized in that: In the step (4), the RNA ligation strand RNA 0-x is designed such that each strand contains 24 bases, with 12 bases at one end connected to the detection strand D0 and 12 bases at the other end connected to the detection strand D x to form a circular DNA nanoswitch.

8. A binary digital encryption operation method based on a DNA nanoswitch according to claim 7, characterized in that: The reading of the operation result in the step (5) is completed by adding the target strand T x after incubation; the target strand T x is designed such that each strand contains 24 bases, with 12 bases at one end connected to the detection strand D0 and 12 bases at the other end connected to the detection strand D x connected.

9. A binary digital encryption operation method based on a DNA nanoswitch according to claim 8, characterized in that: In step (5), the binary digital encryption operation strategy reads the operation result by observing the position of the circular DNA nanoswitch on the gel diagram through agarose gel electrophoresis. The agarose gel electrophoresis conditions are set as follows: agarose concentration of 1.2%, 0.5×TBE buffer, voltage of 60V, and running gel time of 1.5 hours.

10. A binary digital encryption operation method based on a DNA nanoswitch according to claim 9, characterized in that: The target strand T in step (5) x is designed such that each strand contains 24 bases, with 12 bases at one end connected to the detection strand D0 and 12 bases at the other end connected to the detection strand D x for triggering the operation and decryption.

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