A method for building a 9-bit parity check molecular switch circuit

Abstract

The application provides a method for building a 9-bit parity check molecular switch circuit, and the steps are as follows: based on DNA strand displacement technology, the structure and reaction mechanism of the molecular switch circuit are described; the reaction rate and leakage rate are set, and the input signal of the molecular switch circuit and the concentration ratio of the molecular switch at each stage are optimized; the fan-in molecular switch circuit and the fan-out molecular switch circuit are set according to the molecular switch circuit; the 3-bit parity check molecular switch circuit is built based on the fan-in molecular switch circuit and the fan-out molecular switch circuit by using the switch canvas routing strategy, the 9-bit parity check molecular switch circuit is built by using three 3-bit parity check molecular switch circuits with the same structure; and the correctness of the 9-bit parity check molecular switch circuit is verified by using a Simbiology simulation platform. In the construction of a large-scale complex logic circuit, the application has a simple and unified DNA chain structure, a fast reaction speed and good stability, and provides a new scheme for the construction of a modular DNA circuit.

Description

Technical Field

[0001] This invention relates to the technical field of biological logic circuits, and in particular to a method for constructing a 9-bit parity check molecular switch circuit, which utilizes DNA strand substitution technology to construct the 9-bit parity check molecular switch logic circuit. Background Technology

[0002] In recent years, DNA has been widely used to construct biomolecular systems due to its base pair specificity and flexible programming capabilities. Among these, DNA strand substitution technology is considered an ideal mechanism for constructing complex biological network structures. It is often combined with DNA origami and fluorescent labeling techniques to achieve nanoscale self-assembly and digital simulation computing. Over the past few decades, it has been developed and applied in fields such as biomedicine, nonlinear systems, DNA logic computing, and neural networks. For example, DNA nanostructures constructed using DNA strand substitution and DNA origami can be used for intelligent drug targeted delivery to treat diseases. Artificial neural networks based on strand substitution implement XOR operations and full adder operations, and DNA circuits can realize many complex logical functions. Currently, DNA logic circuits based on DNA strand displacement are a hot research direction and have made great progress over the past few decades. Some simple DNA logic gates developed early on are the primitive elements of complex molecular logic circuits, such as AND gates, OR gates, NOT gates, and integrated gates. Later, complex DNA logic circuits, such as encoders, multiplexers, full adders and subtractors, timers, and matrix multiplication circuits, were realized by cascading basic logic gates. These logic circuits are the necessary foundation for the future realization of biological computers, and can effectively elevate bioengineering technology and artificial intelligence to a higher level. Summary of the Invention

[0003] To address the technical challenges of complex circuit structures, slow response speeds, numerous and disorganized participating chains, and lack of modularity when constructing large-scale complex circuits, this invention proposes a method for building a 9-bit parity check molecular switch circuit. A DNA switch circuit model is built using the Simbiology simulation platform in Matlab, elucidating the reaction mechanism of the molecular switch circuit and achieving the expected function, thus verifying its feasibility.

[0004] To achieve the above objectives, the technical solution of the present invention is as follows: A method for constructing a 9-bit parity check molecular switch circuit, comprising the following steps:

[0005] Step 1: Elucidate the structure and reaction mechanism of the molecular switch circuit based on DNA strand substitution technology;

[0006] Step 2: Set the reaction rate and leakage rate, and optimize the input signal of the molecular switch circuit and the concentration ratio of each stage of the molecular switch;

[0007] Step 3: Set up the fan-in molecular switch circuit and the fan-out molecular switch circuit according to the molecular switch circuit;

[0008] Step 4: Using the switch canvas routing strategy, build a 3-bit parity check molecular switch circuit based on the fan-in molecular switch circuit and the fan-out molecular switch circuit. Use four identical 3-bit parity check molecular switch circuits to build a 9-bit parity check molecular switch circuit.

[0009] Step 5: Verify the correctness of the 9-bit parity check molecular switch circuit using the Simbiology simulation platform.

[0010] The molecular switch circuit in step one includes a start switch SW, a downstream switch DS that receives upstream signals, and a fluorescence reporter gate Report; the chemical reaction mechanism of the molecular switch circuit is as follows:

[0011] A+SW→CA+SW(on) (1)

[0012] CA+DS→DS(off)+waste1 (2)

[0013] B+DS(off)→CY+DS(on) (3)

[0014] CY+Report→Report(on)+waste2 (4)

[0015] Among them, reaction (1) is the chain substitution reaction between input signal A and start switch SW, where A and SW are both reactants, and CA and SW(on) are both products. CA represents the generated current signal chain, and SW(on) represents the start switch SW being closed. Reaction (2) is the chain substitution reaction between current signal chain CA and downstream switch DS which is in a blocked state. CA and DS are both reactants, and DS(off) and waste1 are both new products. DS(off) represents the downstream switch DS changing from the original blocked state to the open state, and waste1 is waste. Reaction (3) is the chain substitution reaction between input signal B and switch DS(off), producing products CY and DS(on). CY is the current signal chain, and DS(on) represents the downstream switch DS changing to the closed state. Reaction (4) is the chain substitution reaction between current signal chain CY and fluorescence reporter gate Report. Report(on) is the generated fluorescence signal chain, representing the fluorescence reporter gate Report being closed. This reaction process converts the concentration of signal chain CY into fluorescence intensity output.

[0016] The reaction process is as follows: Input signal A enters from the input terminal and combines with the exposed foothold of start switch SW, thereby generating a current signal chain CA; downstream switch DS consists of S domain and D domain. S domain receives the current signal chain CA transmitted from upstream switch through the exposed foothold, while D domain responds to the input signal B of downstream switch. After the current signal chain CA is replaced by the downstream switch DS chain, the downstream switch DS is switched to the open state, waiting for the arrival of input signal Y; after the downstream switch DS is replaced by the input signal Y, a current signal chain CY is generated, and the downstream switch DS is switched to the closed state DS(on). Finally, the current signal chain CY reacts with the fluorescence reporter gate to generate a fluorescence signal chain that is detected and output.

[0017] In the reaction formula (1), the DNA single-strand structure of the input signal A is as follows:<n1^s1^s2^n2^n3^> The DNA double-stranded structure of the start switch SW is<t2^t1 t2^> [n1^s1^s2^n2^]{n3^*}, the single-stranded DNA structure of the product CA is as follows:<t2^t1t2^n1^s1^s2^n2^> The DNA double-stranded structure of the closed switch SW(on) is [n1^s1^s2^n2^n3^]; in the reaction formula (2), the DNA double-stranded structure of the downstream switch DS is<u3^v3 u4^> [u1^v1^v2^]:[u5^t2^t1 t2^]{n1^*}, the DNA double-stranded structure of the product DS(off) is as follows:<u3^v3 u4^> [u1^v1^v2^]:{u5^}:[t2^t1 t2^n1^]<s1^s2^n2^> In the reaction formula (3), the DNA single-strand structure of the input signal Y is as follows:<u1^v1^v2^u5^> After the input signal B undergoes a strand substitution reaction with DS(off), the single-stranded DNA structure of the product CY is as follows:<u3^v3^u4^u1^v1^v2^> The DNA double-stranded structure of the product DS(on) is [u1^v1^v2^u5^]:[t2^t1 t2^n1^]<s1^s2^n2^> In the reaction formula (4), the DNA double-stranded structure of the fluorescent reporter gate Report is [u1^v3 u4^]{u1^*}, and the DNA double-stranded structure of the product Report(on) is [u1^v3 u4^u1^]{v1^v2^}; where s1, s2, n2, t1, t2, u3, u4, v1, v2, v3 are structural domains, u1, n1, n3, u5 are small fulcrum domains, <> represents the upper strand structure of the DNA strand, [] represents the double-stranded structure of the DNA strand that has been complementary, : represents the double-stranded structure used to connect the two double-stranded structural domains, {} represents the lower strand structure of the DNA strand, ^ is used to mark the upper strand structural domain, and ^* is used to mark the lower strand structural domain;

[0018] In step two, the reaction rate of the molecular switch circuit is set to 1.0E-4nMs. -1 The leakage rate of the switch in the blocked state is set to 1.0E-6nM. -1 s -1 The leakage rate of the switch in the open state is set to 5.0E-6 nMs. -1 The initial concentration of input signal A is set to twice the concentration of the start switch SW. If the upstream switch and the downstream switch are connected in series, the initial concentration ratio of the molecular switches is set to 1.5x:1x, where x = 100nM.

[0019] The fan-in molecular switch circuit includes a 2-fan-in molecular switch circuit, which aggregates two current signals transmitted from the upstream switch into a single current signal. The 2-fan-in molecular switch circuit includes upstream switches DS1 and DS2, and downstream switch SW1. Upstream switches DS1 and DS2 are connected in parallel and then in series with the downstream switch SW1. The reaction equation for the 2-fan-in molecular switch circuit is:

[0020] A1+DS1→CA1+DS1(on) (5)

[0021] A2+DS2→CA1+DS2(on) (6)

[0022] CA1+SW→SW(off) (7)

[0023] B+SW(off)→CS+SW(on) (8)

[0024] In reactions (5) and (6), the input signals A1 and A2 undergo chain substitution reactions with the upstream switches DS1 and DS2 respectively, generating the same current signal chain CA1, and switching the states of the downstream switches DS1 and DS2 to the closed states DS1(on) and DS2(on) respectively. In reaction (7), the current signal chain CA1 undergoes chain substitution reactions with the downstream switch SW, and the current signal chain CA1 activates the downstream switch SW. The state of the downstream switch SW changes from the original blocked state to the open state SW1(off), exposing the middle small fulcrum to wait for the arrival of the input signal B. In reaction (8), the input signal B undergoes chain substitution reactions with the downstream switch SW(off) to generate a current signal chain CS, and the state of the downstream switch SW changes to the closed state SW(on).

[0025] The fan-out molecular switch circuit includes a 2-fan-out molecular switch circuit, which includes an upstream switch SW and two downstream switches DS1 and DS2 connected in parallel. The upstream switch SW is connected in series with the parallel downstream switches DS1 and DS2.

[0026] The reaction equation for the 2-fan-out molecular switch circuit is:

[0027] A+SW→CA+SW(on) (9)

[0028] CA+DS1→DS1(off)+waste1 (10)

[0029] CA+DS2→DS2(off)+waste2 (11)

[0030] B1+DS1(off)→CB+DS1(on) (12)

[0031] B2+DS2(off)→CB+DS2(on)(13)

[0032] In reaction (9), the input signal A is replaced by the upstream switch SW chain to generate the current signal chain CA, and the upstream switch SW is converted to the closed state SW(on); in reaction (10)-(11), the two fan-out current signal chains CA are simultaneously transmitted to the downstream switches DS1 and DS2 and activate them, and the states of the downstream switches DS1 and DS2 are converted to the open states DS1(off) and DS2(off) respectively; in reaction (12)-(13), the input signals B1 and B2 react with the exposed DS1(off) and DS2(off) with the middle small support point respectively to generate the same current signal chain CB, double chain DS1(on) and double chain DS2(on), and the downstream switches DS1 and DS2 are converted from the open state to the closed state. At this time, the current signal chain CB is the output signal of the 2-fan-out molecular switch circuit.

[0033] In the reaction equations (6) and (7) of the 2-fan-in molecular switch circuit, the DNA single-strand structures of the input signals A1 and A2 are designed as follows:<n1^s1^s2^n2^n3^> and<n1^s1^s2^n2^n4^> The DNA double-stranded structures of upstream switches DS1 and DS2 are respectively<t2^t1 t2^> [n1^s1^s2^n2^]{n3^*} and<t2^t1 t2^> [n1^s1^s2^n2^]{n4^*}, the single-stranded DNA structure of the product CA1 is as follows:<t2^t1 t2^n1^s1^s2^n2^> The DNA double-stranded structure of the downstream switch SW1 in reaction (8) is as follows:<u2^v3 u3^> [u1^v1^v2^]:[u4^t2^t1 t2^]{n1^*};The single-stranded DNA structure of the input signal strand Y in reaction (9) is<u1^v1^v2^u4^> The generated current signal chain CS is a single-stranded DNA.<u2^v3 u3^u1^v1^v2^> ; where t1, t2, s1, s2, n2, u1, u2, u3, v1, v2, v3 are structural domains, n1, n3, n4, u4 are small fulcrum domains, <> represents the upper strand structure of the DNA strand, [] represents the double-stranded structure of the DNA strand that has been complementary, : represents the double-stranded structural domain used to connect the two double-stranded structural domains, {} represents the lower strand structure of the DNA strand, ^ is used to mark the upper strand structural domain, and ^* is used to mark the lower strand structural domain.

[0034] The initial concentrations of upstream switches DS1 and DS2 are both set to 1.5x, the initial concentration of downstream switch SW1 is set to 1x, the initial concentrations of all input signals are set to twice the initial concentration of the molecular switch, and the initial concentration of input signal X1 of upstream switch DS1 is set to 3x.

[0035] In the reaction equation (10) of the 2-fan-out molecular switch circuit, the DNA single-strand structure of the input signal A is as follows:<n1^s1^s2^n2^n3^> The DNA double-stranded structure of the upstream switch SW is<t2^t1 t2^> [n1^s1^s2^n2^]{n3^*}, the single-stranded DNA structure of the product CA is as follows:<t2^t1 t2^n1^s1^s2^n2^> The DNA double-stranded structures of downstream switch DS1 and downstream switch DS2 in the reaction formulas (11)-(12) are respectively<u2^v3 u3^> [u1^v1^v2^]:[u4^t2^t1 t2^]{n1^*} and<u2^v3u3^> [u1^v1^v2^]:[u5^t2^t1 t2^]{n1^*};The DNA single-strand structures of the input signals Y1 and Y2 in the reaction formulas (13)-(14) are respectively<u1^v1^v2^u4^> and<u1^v1^v2^u5^> The generated current signal chain CS is<u2^v3 u3^u1^v1^v2^> ; where s1, s2, n2, u1, u3, t1, t2, v1, v2, v3 are structural domains, n1, n3, u4, u5 are small fulcrum domains, <> represents the upper strand structure of the DNA strand, [] represents the double-stranded structure of the DNA strand that has been complementary, : represents the double-stranded structural domain used to connect the two double-stranded structural domains, {} represents the lower strand structure of the DNA strand, ^ is used to mark the upper strand structural domain, and ^* is used to mark the lower strand structural domain.

[0036] The initial concentration of the upstream switch SW2 is set to 1.5x, the initial concentration of the downstream switches DS3 and DS4 is set to 1x, the initial concentration of all input signals is set to twice the initial concentration of the molecular switch, and the initial concentration of the input signal A corresponding to the upstream switch SW2 is set to 3x.

[0037] The method for constructing the 3-bit parity check molecular switch circuit is as follows: a 3-bit parity check switch circuit is constructed using a switch canvas strategy; a DNA switch circuit with two layers of 10 molecular switches is designed based on the DNA strand substitution reaction.

[0038] The 9-bit parity check molecular switch circuit includes four identical 3-bit parity check molecular switch circuits. The parity of the three identical 3-bit parity check molecular switch circuits is detected by detecting the parity of the three sets of binary codes respectively. The output current signal chain is used as the input signal of the fourth parity check module to perform the final parity check and finally obtain the check result.

[0039] The 3-bit parity check molecular switch circuit includes input start switches SW1 and SW2, intermediate downstream switches DS3-DS6, and output downstream switches DS7-DS6. 10And two fluorescent reporter gates, Report0 and Report1, whose DNA strand substitution reactions include:

[0040] The process of input signal A' activating start switch SW1 is as follows:

[0041] A'+SW1→CSA'+SW1(on)(14)

[0042] The reaction of the 2-fan-out molecular switch circuit is:

[0043] CSA'+DS3→DS3(off)+waste1 (15)

[0044] CSA'+DS4→DS4(off)+waste2 (16)

[0045] In reaction (14), after the input signal A' is chain-replaced with the start switch SW1, a current signal chain CSA' is generated, and the start switch SW1 is turned into the closed state SW1(on); in reaction (15)-(16), the current signal chain CSA' is fanned out twice and then transmitted to the downstream switch DS3 and downstream switch DS4 which are in the blocked state. The downstream switch DS3 and downstream switch DS4 are turned into the open state DS3(off) and DS4(off) respectively, with the products waste1 and waste2 being waste materials;

[0046] The process by which input signal B' and input signal B activate downstream switches DS3 and DS4 respectively and output current signal chains is as follows:

[0047] B'+DS3(off)→CSB'+DS3(on) (17)

[0048] B+DS4(off)→CSB+DS4(on) (18)

[0049] In reaction formulas (17)-(18), input signal B' and input signal B react chemically with downstream switches DS3(off) and DS4(off) which are in the off state, respectively, generating two current signal chains CSB' and CSB, which are respectively transmitted to downstream switches DS7 and DS8. 10 At downstream switches DS8 and DS9, and change downstream switches DS3 and DS4 from open to closed states DS3(on) and DS4(on);

[0050] Input signal A activates start switch SW2 as follows:

[0051] A+SW2→CSA+SW2(on)(19)

[0052] The reaction of the 2-fan-out molecular switch circuit is:

[0053] CSA+DS5→DS5(off)+waste3(20)

[0054] CSA+DS6→DS6(off)+waste4(21)

[0055] In reaction (19), after the input signal A is chain-replaced with the start switch SW2, a current signal chain CSA is generated, and the start switch SW2 is turned into the closed state SW2(on); in reactions (20)-(21), the current signal chain CSA is fanned out twice and then transmitted to the downstream switches DS5 and DS6 which are in the blocked state. The downstream switches DS5 and DS6 are turned into the open state DS3(off) and DS4(off) with the middle exposed small fulcrum. The products waste1 and waste2 are waste materials;

[0056] The process by which input signal B' and input signal B react chemically with downstream switches DS5(off) and DS6(off), which are in the off state, is as follows:

[0057] B'+DS5(off)→CSB'+DS5(on) (22)

[0058] B+DS6(off)→CSB+DS6(on) (23)

[0059] In reaction formulas (22)-(23), input signal B' and input signal B react chemically with downstream switches DS5(off) and DS6(off) which are in the off state, respectively, to generate two current signal chains CSB' and CSB, which are transmitted to downstream switches DS8 and DS9 and downstream switches DS7 and DS8 respectively. 10 At this point, the downstream switches DS5 and DS6 are changed from the open state to the closed state DS5(on) and DS6(on);

[0060] The upstream current signal chains CSB' and CSB activate the downstream switches DS7 and DS, respectively. 10 The reaction process with DS8 and DS9 is as follows:

[0061] CSB'+DS7→DS7(off)+waste5 (24)

[0062] CSB'+DS 10 →DS 10 (off)+waste6 (25)

[0063] CSB+DS8→DS8(off)+waste7 (26)

[0064] CSB+DS9→DS9(off)+waste8 (27)

[0065] Among them, waste5, waste6, waste7, and waste8 are waste materials, and the activated downstream switches DS7, DS8, DS9, and DS... 10 The previously blocked states were respectively switched to open states: DS7(off), DS8(off), DS9(off), DS 10 (off), exposing the small central pivot point to await the arrival of input signals C and C';

[0066] Input signals C', C', C', and C' are respectively connected to downstream switches DS7(off), DS8(off), DS9(off), and DS'(off) which are in the off state. 10 The process by which (off) produces a chemical reaction is as follows:

[0067] C'+DS7(off)→Σ O +DS7(on) (28)

[0068] C+DS8(off)→Σ O +DS8(on) (29)

[0069] C'+DS9(off)→Σ E +DS9(on) (30)

[0070] C+DS 10 (off)→Σ E +DS 10 (on) (31)

[0071] Among them, the downstream switches DS7 to DS 10 The switches transition from the open state to the closed state respectively: DS7(on), DS8(on), DS9(on), DS 10 (on), Σ O , Σ E For the two generated current signal chains, current signal chain Σ O , Σ E It is transmitted to the fluorescence reporting gates Report0 and Report1;

[0072] Current signal chain Σ O With the fluorescence report gate Report0 and the current signal chain Σ E The reaction processes with the fluorescent reporter gate Report1 are as follows:

[0073] Σ O +Report0→Report0(on)+waste9 (32)

[0074] Σ E +Report1→Report1(on)+waste 10 (33)

[0075] The closure of the two fluorescent reporter gates, Report0 and Report1, generates two fluorescent signal chains, Report0(on) and Report1(on), respectively, producing waste9 and waste. 10 Waste material;

[0076] The DNA chemical reaction rate is set to 1.0E-4 nMs in the 3-bit parity check molecular switch circuit. -1 The leakage rate of the switch in the blocked state is set to 1.0E-6 nMs. -1 The leakage rate of the switch in the open state is set to 5.0E-6 nMs. -1 The initial concentration of the molecular switch is set to 1xnM, where x represents 100. The initial concentrations of input signals A, B, and C are set to 2xnM. The concentration ratio of each stage of the molecular switch is set to 4:1.5:1.

[0077] The chemical reaction equation for the fourth parity check module is:

[0078] Σ E1 +SW1→ΣC E1 +SW1(on) (34)

[0079] ΣC E1 +DS3→DS3(off)+waste1 (35)

[0080] ΣC E1 +DS4→DS4(off)+waste2 (36)

[0081] Σ E2 +DS3(off)→ΣC E2 +DS3(on) (37)

[0082] Σ o2 +DS4(off)→ΣC O2 +DS4(on) (38)

[0083] ΣC E2 +DS7→DS7(off)+waste3 (39)

[0084] ΣC E2 +DS 10 →DS 10 (off)+waste4 (40)

[0085] ΣCO2 +DS8→DS8(off)+waste5 (41)

[0086] ΣC O2 +DS9→DS9(off)+waste6 (42)

[0087] Σ O3 +DS7(off)→CY O +DS7(on) (43)

[0088] Σ E3 +DS8(off)→CY O +DS8(on) (44)

[0089] Σ O1 +SW2→ΣC O2 +SW2(on) (45)

[0090] ΣC O2 +DS5→DS5(off)+waste7 (46)

[0091] ΣC O2 +DS6→DS6(off)+waste8 (47)

[0092] Σ E2 +DS5(off)→ΣC E2 +DS5(on) (48)

[0093] Σ o2 +DS6(off)→ΣC O2 +DS5(on) (49)

[0094] ΣC E2 +DS8→DS8(off)+waste9 (50)

[0095] ΣC E2 +DS9→DS9(off)+waste 10 (51)

[0096] ΣC O2 +DS7→DS7(off)+wast 11 (52)

[0097] ΣC O2 +DS 10 →DS 10 (off)+waste 12 (53)

[0098] Σ O3 +DS9(off)→CYE +DS9(on) (54)

[0099] Σ E3 +DS 10 (off)→CY E +DS 10 (on) (55)

[0100] Σ O +Report0→Report0(on)+waste 13 (56)

[0101] Σ E +Report1→Report1(on)+waste 14 (57)

[0102] Among them, the generated waste1-waste 14 As waste, the reaction process of reaction (34)-(57) is as follows: the output chain Σ from the first three 3-bit parity check molecular switch circuits is converted into waste. O1 , Σ E1 , Σ O2 , Σ E2 , Σ O3 , Σ E3 As the input signal chain for each molecular switch; when the output chain Σ O1 , Σ E1 Upon arrival, upstream switches SW1 and SW2 close, and the output current signal chain ΣC is activated. O1 , ΣC E1 Activate downstream switches DS3-DS6, and wait for the output chain Σ. O2 , Σ E2 The input; when the output chain Σ O2 , Σ E2 After input, the corresponding downstream switches DS3-DS6 turn to the closed state, and the output current signal chain ΣC O2 , ΣC E2 ; Current signal chain ΣC O2 , ΣC E2 Activate downstream switch DS7-DS 10 Waiting for the output chain Σ O3 , Σ E3 The input; when the output chain Σ O3 , Σ E3 After the input, the downstream switch DS7-DS 10 Switching to a closed state, the output current signal chain Σ O , Σ EThe signal is transmitted to the fluorescence reporter gates Report0 and Report1. After reacting with the fluorescence reporter gates, two fluorescence signal chains, Report0(on) and Report1(on), are output as the final verification results. One of the two fluorescence chains reaches dynamic equilibrium after 1000 seconds, while the other outputs a low fluorescence signal of less than 10 nM.

[0103] The beneficial effects of this invention are:

[0104] 1. This invention utilizes DNA strand substitution technology to construct a molecular switch circuit, designs a molecular switch chain structure, and optimizes the concentration ratio of each stage of the molecular switch, verifying that the molecular switch has AND, OR, and fan-in / fan-out cascade functions.

[0105] 2. This invention combines a switch canvas routing strategy with DNA strand replacement technology, and successfully uses a DNA switch circuit to implement 9-bit parity check operation.

[0106] 3. The DNA switch circuit of the present invention uses a simple molecular switch sequence to replace the complex logic gate construction process, which improves the reaction speed and effectively simplifies the circuit scale.

[0107] 4. This invention constructs a parity check DNA switch circuit capable of processing 512 groups of 9-bit binary codes, verifying the ability of DNA switch circuits to handle large-scale complex circuits and promoting the modularization of DNA computing. Attached Figure Description

[0108] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0109] Figure 1 This is a flowchart illustrating the implementation of the 9-bit parity check molecular switch circuit of the present invention.

[0110] Figure 2 This is a block diagram of the 9-bit parity check molecular switch circuit of the present invention.

[0111] Figure 3 This is a schematic diagram of the switch canvas structure of the 3-bit parity check switch circuit of the present invention.

[0112] Figure 4 This is a schematic diagram of the 3-bit parity check molecular switch circuit of the present invention.

[0113] Figure 5 This is a schematic diagram of the 3-bit parity check molecular switch circuit model of the present invention.

[0114] Figure 6 The simulation results of the 3-bit parity check switch circuit of the present invention are shown in Figure (a), where ABC = 000 and Σ O Σ E =10, (b) is ABC=001 and Σ O Σ E =01, (c) is ABC =010 and Σ O Σ E =01, (d) is ABC =011 and Σ O Σ E =10, (e) is ABC = 100 and Σ O Σ E =01, (f) is ABC = 101 and Σ O Σ E =10, (g) is ABC = 110 and Σ O Σ E =10, (h) is ABC = 111 and Σ O Σ E =01.

[0115] Figure 7 The simulation results of the 9-bit parity check switch circuit of the present invention are shown in the figure, where (a) is GHI-DEF-ABC=001-010-111 and Σ O Σ E =01, (b) is GHI-DEF-ABC=001-111-011 and Σ O Σ E =10, (c) is GHI-DEF-ABC=010-011-000 and Σ O Σ E =01, (d) is GHI-DEF-ABC=011-100-111 and Σ O Σ E =10, (e) is GHI-DEF-ABC=100-010-001 and Σ O Σ E =01, (f) is GHI-DEF-ABC=101-011-011 and Σ O Σ E =10, (g) is GHI-DEF-ABC=110-101-010 and Σ O Σ E =01, (h) is GHI-DEF-ABC=111-100-100 and Σ O Σ E =10, (I) is GHI-DEF-ABC=111-111-111 and Σ O ΣE =01. Detailed Implementation

[0116] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0117] like Figure 1 As shown, a method for constructing a 9-bit parity check molecular switch circuit is presented. First, the basic structure of the molecular switch circuit is constructed based on DNA strand substitution technology, verifying that the molecular switch possesses series-parallel connection and fan-in / fan-out functionality. Second, a 9-bit parity check DNA switch circuit is constructed, consisting of four sets of parity check DNA switch circuits. Each of these four sets of DNA switch circuits comprises two layers of 10 molecular switch circuits. The input signal concentration and the concentration ratio of each molecular switch stage in each set of DNA switch circuits are rationally optimized to achieve optimal output. Each DNA switch circuit can receive three input signal chains. The first three sets of parity check switch circuits each output a 3-bit binary code parity check result. The last parity check DNA switch circuit uses the first three parity check results as input signals to complete the final 9-bit binary code parity check. Finally, simulation analysis was performed using the Simbiology platform to verify its feasibility. The specific implementation steps are as follows:

[0118] Step 1: Construct a molecular switch circuit structure based on the DNA strand substitution reaction mechanism. The structure of the molecular switch circuit includes a start switch SW, a downstream switch DS that receives upstream signals, and a fluorescence reporter gate Report. The chemical reaction mechanism of the molecular switch circuit is as follows:

[0119] A+SW→CA+SW(on) (1)

[0120] CA+DS→DS(off)+waste1 (2)

[0121] B+DS(off)→CY+DS(on) (3)

[0122] CY+Report→Report(on)+waste2 (4)

[0123] In this reaction, reaction (1) is the chain substitution reaction between input signal A and start switch SW, where A and SW are reactants, and CA and SW(on) are products. CA represents the generated current signal chain, and SW(on) represents the closing of start switch SW. Reaction (2) is the chain substitution reaction between current signal chain CA and downstream switch DS, which is in a blocked state. CA and DS are reactants, and DS(off) and waste1 are new products. DS(off) represents the downstream switch DS changing from a blocked state to an open state, and waste1 is waste. Reaction (3) is the chain substitution reaction between input signal B and switch DS(off), generating CY and DS(on). CY is the current signal chain, and DS(on) represents the downstream switch DS changing to a closed state. Reaction (4) is the chain substitution reaction between current signal chain CY and fluorescence reporter gate Report. Report(on) is the generated fluorescence signal chain, representing the closing of fluorescence reporter gate Report. This reaction process converts the concentration of signal chain CY into fluorescence intensity output.

[0124] The specific process of the molecular switch circuit reaction is as follows: Input signal A enters from the input terminal and combines with the exposed foothold of the start switch SW, thereby generating a current signal chain CA; the downstream switch DS consists of two domains, namely the S domain and the D domain. The S domain receives the current signal chain CA transmitted from the upstream switch through the exposed foothold, while the D domain responds to the input signal B of the downstream switch. After the current signal chain CA is replaced by the DS chain, the DS state is changed to the open state, waiting for the arrival of the input signal Y; after the downstream switch DS is replaced by the input signal Y, a current signal chain CY is generated, and the DS is changed to the closed state DS(on). Finally, after CY reacts with the fluorescence reporter gate, a fluorescence signal chain is generated and detected and output. When there is no input signal, the start switch will not be activated and closed, the current signal chain will not be generated, the downstream switch connected in series will not be closed, and the fluorescence reporter gate will not output a fluorescence signal.

[0125] In reaction (1), the DNA single-strand structure of the input signal A is designed as follows:<n1^s1^s2^n2^n3^> The DNA double-stranded structure of the start switch SW is designed as follows:<t2^t1 t2^> [n1^s1^s2^n2^]{n3^*}, the single-stranded DNA structure of the product CA is as follows:<t2^t1 t2^n1^s1^s2^n2^> The DNA double-stranded structure of the closed switch SW(on) is [n1^s1^s2^n2^n3^]; in the reaction formula (2), the DNA double-stranded structure of the downstream switch DS is<u3^v3 u4^> [u1^v1^v2^]:[u5^t2^t1t2^]{n1^*}, the DNA double-stranded structure of the product DS(off) is as follows:<u3^v3 u4^> [u1^v1^v2^]:{u5^}:[t2^t1 t2^n1^]<s1^s2^n2^> In the reaction formula (3), the DNA single-strand structure of the input signal Y is as follows:<u1^v1^v2^u5^> After the input signal B undergoes a strand substitution reaction with DS(off), the single-stranded DNA structure of the product CY is as follows:<u3^v3^u4^u1^v1^v2^> The DNA double-stranded structure of the product DS(on) is [u1^v1^v2^u5^]:[t2^t1 t2^n1^]<s1^s2^n2^> In the reaction formula (4), the DNA double-stranded structure of the fluorescent reporter gate Report is [u1^v3 u4^]{u1^*}, and the DNA double-stranded structure of the product Report(on) is [u1^v3 u4^u1^]{v1^v2^}; where s1, s2, n2, t1, t2, u3, u4, v1, v2, v3 are structural domains, u1, n1, n3, u5 are small fulcrum domains, <> represents the upper strand structure of the DNA strand, [] represents the double-stranded structure of the DNA strand that has been complementary, : represents the double-stranded structure used to connect the two double-stranded structural domains, {} represents the lower strand structure of the DNA strand, ^ is used to mark the upper strand structural domain, and ^* is used to mark the lower strand structural domain.

[0126] Step 2: Set the reaction rate and leakage rate between DNA strands, and optimize the concentration ratio of each input signal and the molecular switch. The reaction rate of the molecular switch circuit is set to 1.0E-4 nMs. -1 The leakage rate of the switch in the blocked state is set to 1.0E-6 nMs. -1 The leakage rate of the switch in the open state is set to 5.0E-6 nMs. -1 The initial concentration of input signal A is set to twice the concentration of the start switch SW. If the upstream switch and the downstream switch are connected in series, the initial concentration ratio of the molecular switches is set to 1.5x:1x, where x = 100nM.

[0127] Step 3: Set up the fan-in molecular switch circuit, including a 2-fan-in molecular switch circuit. The fan-in function of the molecular switch circuit aggregates several current signals transmitted from the upstream switch into a single current signal. The 2-fan-in molecular switch circuit includes upstream switches DS1 and DS2 and downstream switch SW1. Upstream switches DS1 and DS2 are connected in parallel and then in series with downstream switch SW1. The reaction equation of the 2-fan-in molecular switch circuit is:

[0128] A1+DS1→CA1+DS1(on) (5)

[0129] A2+DS2→CA1+DS2(on) (6)

[0130] CA1+SW→SW(off) (7)

[0131] B+SW(off)→CS+SW(on) (8)

[0132] In reactions (5)-(6), the input signals A1 and A2 undergo chain substitution reactions with the upstream switches DS1 or DS2 respectively, generating the same current signal chain CA1, and switching the states of switches DS1 and DS2 to the closed states DS1(on) and DS2(on). In reaction (7), the current signal chain CA1 undergoes chain substitution reactions with the downstream switch SW, and the current signal chain CA1 activates the downstream switch SW. The state of SW changes from the original blocked state to the open state SW1(off), exposing the middle small fulcrum to wait for the arrival of the input signal B. In reaction (8), the input signal B undergoes chain substitution reactions with the downstream switch SW(off), generating a current signal chain CS, and the state of the downstream switch SW changes to the closed state SW(on).

[0133] In the 2-fan-in molecular switch circuit, the DNA single-strand structures of the input signals A1 and A2 are designed as follows:<n1^s1^s2^n2^n3^> and<n1^s1^s2^n2^n4^> The DNA double-stranded structures of upstream switches DS1 and DS2 are designed as follows:<t2^t1 t2^> [n1^s1^s2^n2^]{n3^*} and<t2^t1 t2^> [n1^s1^s2^n2^]{n4^*}, the single-stranded DNA structure of the product CA1 is as follows:<t2^t1 t2^n1^s1^s2^n2^> The DNA double-stranded structure of the downstream switch SW1 in reaction (8) is as follows:<u2^v3 u3^> [u1^v1^v2^]:[u4^t2^t1 t2^]{n1^*};The single-stranded DNA structure of the input signal strand Y in reaction (9) is<u1^v1^v2^u4^> The generated current signal chain CS is a single-stranded DNA.<u2^v3 u3^u1^v1^v2^> ; where t1, t2, s1, s2, n2, u1, u2, u3, v1, v2, v3 are structural domains, n1, n3, n4, u4 are small fulcrum domains, <> represents the upper strand of the DNA strand, [] represents the double-stranded structure of the DNA strand that has been complementary, : represents the double-stranded structural domain used to connect the two parts of the double-stranded structural domain, {} represents the lower strand of the DNA strand, ^ is used to mark the upper strand structural domain, and ^* is used to mark the lower strand structural domain.

[0134] The initial concentrations of upstream switches DS1 and DS2 are set to 1.5x, the initial concentration of downstream switch SW1 is set to 1x, the initial concentrations of all input signals are set to twice the initial concentration of the molecular switch, and the initial concentration of input signal X1 of upstream switch DS1 needs to be set to 3x.

[0135] The fan-out molecular switch circuit includes a 2-fan-out molecular switch circuit, which comprises an upstream switch SW and two downstream switches DS1 and DS2 connected in parallel. The upstream switch SW and the parallel downstream switches DS1 and DS2 are connected in series. The reaction equation of the 2-fan-out molecular switch circuit is:

[0136] A+SW→CA+SW(on) (9)

[0137] CA+DS1→DS1(off)+waste1 (10)

[0138] CA+DS2→DS2(off)+waste2 (11)

[0139] B1+DS1(off)→CB+DS1(on) (12)

[0140] B2+DS2(off)→CB+DS2(on) (13)

[0141] In reaction (9), the input signal A is replaced by the upstream switch SW chain to generate the current signal chain CA, and the upstream switch SW is converted to the closed state SW(on); in reactions (10)-(11), the two fan-out current signal chains CA are simultaneously transmitted to the downstream switches DS1 and DS2 and activate them, and the states of the downstream switches DS1 and DS2 are changed to the open states DS1(off) and DS2(off); in reactions (12)-(13), the input signals B1 and B2 react chemically with the exposed DS1(off) and DS2(off) with the middle small fulcrum to generate the same current signal chain CB, double-stranded DS1(on) and double-stranded DS2(on), and the downstream switches DS1 and DS2 are changed from the open state to the closed state. At this time, the current signal chain CB is the output signal of the 2-fan-out DNA switch circuit.

[0142] In the reaction equation (10) of the 2-fan-out molecular switch circuit, the DNA single-strand structure of the input signal A is:<n1^s1^s2^n2^n3^> The DNA double-stranded structure of the upstream switch SW is<t2^t1 t2^> [n1^s1^s2^n2^]{n3^*}, the single-stranded DNA structure of the product CA is as follows:<t2^t1 t2^n1^s1^s2^n2^> The DNA double-stranded structures of the downstream switches DS1 and DS2 in the reaction formulas (11)-(12) are respectively<u2^v3 u3^> [u1^v1^v2^]:[u4^t2^t1 t2^]{n1^*} and<u2^v3 u3^> [u1^v1^v2^]:[u5^t2^t1 t2^]{n1^*};The DNA single-strand structures of the input signals Y1 and Y2 in reaction formulas (13)-(14) are respectively<u1^v1^v2^u4^> and<u1^v1^v2^u5^> The generated current signal chain CS is<u2^v3 u3^u1^v1^v2^> Among them, s1, s2, n2, u1, u3, t1, t2, v1, v2, v3 are structural domains, and n1, n3, u4, u5 are small pivot domains.

[0143] The initial concentration of upstream switch SW2 is set to 1.5x, the initial concentration of downstream switches DS3 and DS4 is set to 1x, and the initial concentration of all input signals is set to twice the initial concentration of the molecular switch. The initial concentration of input signal A of upstream switch SW2 should be set to 3x.

[0144] Step 4: Using the switch canvas strategy, build a 9-bit parity check molecular switch circuit based on the fan-in molecular switch circuit and the fan-out molecular switch circuit.

[0145] like Figure 2 As shown, the 9-bit parity check molecular switch circuit mainly consists of four 3-bit parity check molecular switch circuits. First, a 2-layer, 10-switch-circuit 3-bit parity check molecular switch circuit is designed using a switch canvas strategy, 2-fan-out molecular switch circuits, and 2-fan-in molecular switch circuits. Second, a 9-bit parity check switch circuit is constructed. The 9-bit parity checker consists of four identical 3-bit parity check molecular switch circuits. The 9-bit parity checker mainly uses three identical 3-bit parity check molecular switch circuits to detect the parity of three sets of binary codes respectively. The output of six current signal chains serves as the input signal for the fourth parity check circuit for a final parity check, ultimately obtaining the check result.

[0146] The specific implementation process of constructing a 9-bit parity check molecular circuit is as follows:

[0147] First, a 3-bit parity check switch circuit is constructed using a switch canvas strategy. For example... Figure 3 As shown, the switch canvas strategy is a programmable switch canvas with complementary switch pairs. It combines a 3-bit parity truth table with the complementary switch pairs of the switch canvas to draw a 3-bit parity check switch circuit. Each horizontal switch A, B, C on the switch canvas corresponds to a logic "1" input, and each vertical switch A', B', C' corresponds to a logic "0" input. O Represents the odd parity output, Σ E This represents the even parity output. All input combinations with an output value of "1" in the truth table are mapped to solid lines on the canvas, and the remaining input combinations are mapped to dashed lines. After combining and rearranging the solid lines, a 3-bit parity check switch circuit with 2 layers and 10 molecular switches is obtained. Here, when the odd parity bit Σo is "1", it indicates that the number of "1"s in the output data is even; when the odd parity bit Σo is "0", it indicates that the number of "1"s in the output data is odd. Even parity bit Σo E The result is exactly the opposite of Σo, if the even parity bit Σ E If the parity is "1", then the number of 1s in the data calculation is odd. If the parity digit Σ is even... E A value of "0" means that the number of 1s in the data is even.

[0148] A DNA circuit with two layers and 10 molecular switches, designed using a switch canvas routing strategy and DNA strand displacement reaction, consists of input start switches SW1 and SW2, intermediate downstream switches DS3-DS6, and output downstream switches DS7-DS6. 10 It consists of two fluorescence reporting gates, Report0 and Report1. Figure 4The diagram shows the chain structure of a 3-bit parity check molecular switch circuit. The DNA chain structures for the start switches SW1 and SW2 are designed as follows:<r2^r1 r2^> [m1^a1^a2^m2^]{m3^*} and<r4^r3r4^> [m4^a1^a2^m5^]{m6^*}, the DNA strand structures of downstream switches DS3-DS6 are respectively<e3^b3e4^> [e1^b1^b2^]:[e2^r2^r1 r2^]{m1^*}、<g3^c1 g4^> [g1^b1^b2^]:[e5^r2^r1 r2^]{m1^*}、<g3^c1 g4^> [g1^b1^b2^]:[e6^r4^r3 r4^]{m4^*}、<e3^b3 e4^> [e1^b1^b2^]:[e7^r4^r3r4^]{m4^*};Downstream switch DS7-DS 10 The DNA strand structures are respectively<g2^b4 g2^> [e9^b5^b6^]:[e8^e3^b3e4^]{e1^*}、<g2^b4 g2^> [e9^b5^b6^]:[g5^g3^c1 g4^]{g1^*}、<g7^b9 g7^> [e10^b7^b8^]:[g6^g3^c1 g4^]{g1^*}、<g7^b9 g7^> [e10^b7^b8^]:[g8^e3^b3 e4^]{e1^*}. Where m1, m3, m4, m6, e1, g1, e6, e7, e8, g5, g6, and g8 are small pivot domains; m2, r1, r2, b1, b2, b3, b4, g1, and g2 are structural domains; <> represents the upper strand of the DNA strand; [] represents the double-stranded structure of the DNA strand that has been complementary; : represents the domain used to connect two double-stranded structures; {} represents the lower strand of the DNA strand; ^ is used to mark the upper strand domain; and ^* is used to mark the lower strand domain.

[0149] The structure of a 3-bit parity check molecular switch circuit is as follows: Figure 2 As shown, the chain substitution reaction process is as follows:

[0150] A'+SW1→CSA'+SW1(on) (14)

[0151] CSA'+DS3→DS3(off)+waste1 (15)

[0152] CSA'+DS4→DS4(off)+waste2 (16)

[0153] In this process, reaction (14) is the process by which the input signal A' activates the start switch SW1, and the product CSA' is transmitted to the downstream switches DS3 and DS4. Reactions (15)-(16) are two-fan-out DNA switch circuits. After the current signal chain CSA' fans out two strands, it activates the downstream switches DS3 and DS4 respectively. After activation, the downstream switches DS3 and DS4 change from the original blocked state to the open state, exposing the middle small fulcrum to wait for the arrival of the input signals B and B'.

[0154] B'+DS3(off)→CSB'+DS3(on) (17)

[0155] B+DS4(off)→CSB+DS4(on) (18)

[0156] Among them, reaction formulas (17) to (18) are the process by which input signals B' and B activate downstream switches DS3 and DS4 and output current signal chains. The input signals B' and B react chemically with the downstream switches DS3(off) and DS4(off) which are in the off state, respectively. The two current signal chains CSB' and CSB generated by the reaction are transmitted to the downstream switches DS7 and DS8 respectively. 10 At downstream switches DS8 and DS9, and at downstream switches DS3 and DS4, change from the open state to the closed state.

[0157] A+SW2→CSA+SW1(on) (19)

[0158] CSA+DS5→DS5(off)+waste3 (20)

[0159] CSA+DS6→DS6(off)+waste4 (21)

[0160] In this process, reaction (19) is the process by which input signal A activates the start switch SW2, and the product CSA is passed to the downstream switch. Reactions (20)-(21) are two fan-out DNA switch circuits. After the current signal chain CSA fans out two strands, it activates molecular switches DS5 and DS6 respectively. After activation, molecular switches DS5 and DS6 change from the original blocked state to the open state, exposing the small fulcrum in the middle to wait for the arrival of input signal chains B and B'.

[0161] B'+DS5(off)→CSB'+DS5(on) (22)

[0162] B+DS6(off)→CSB+DS6(on) (23)

[0163] Among them, reaction formulas (22)-(23) are the process of input signals B' and B reacting with downstream switches DS5(off) and DS6(off) which are in the off state after activation. The two current signal chains CSB' and CSB generated by the reaction are transmitted to downstream switches DS8 and DS9 and downstream switches DS7 and DS8 respectively. 10 At this point, the molecular switches DS5 and DS6 are switched from the open state to the closed state.

[0164] CSB'+DS7→DS7(off)+waste5 (24)

[0165] CSB'+DS 10 →DS 10 (off)+waste6 (25)

[0166] CSB+DS8→DS8(off)+waste7 (26)

[0167] CSB+DS9→DS9(off)+waste8 (27)

[0168] Among them, reaction formulas (24)-(27) are the upstream current signal chain CSB', CSB activates the downstream switches DS7 and DS8. 10 The reaction process with DS8 and DS9. The downstream switches DS9 and DS9 after activation. 10 and DS 11 It changes from the original blocked state to the open state, exposing the small fulcrum in the middle to wait for the arrival of the input signal chain C and C'.

[0169] C'+DS7(off)→Σ O +DS7(on) (28)

[0170] C+DS8(off)→Σ O +DS8(on) (29)

[0171] C'+DS9(off)→Σ E +DS9(on) (30)

[0172] C+DS 10 (off)→Σ E +DS 10 (on) (31)

[0173] Among them, reaction formulas (28)-(31) are the input signals C and C' and the DS7 and DS in the disconnected state. 10 The process of chemical reaction between DS7 and DS8, DS9. After the reaction, DS7 to DS9 on the right side... 10 The molecular switch changes from an open state to a closed state, ΣO , Σ E These are the two newly generated current signal chains, which are then passed to the fluorescent reporting gates Report0 and Report1.

[0174] Σ O +Report0→Report0(on)+waste9 (32)

[0175] Σ E +Report1→Report1(on)+waste 10 (33)

[0176] Among them, reaction formulas (32)-(33) are two current signal chains Σ O , Σ E The reaction process with the fluorescent reporter gates Report0 and Report1: After the two fluorescent reporter gates Report0 and Report1 are closed, two fluorescent signal chains, Report0(on) and Report1(on), are generated respectively.

[0177] A schematic diagram of a 3-bit parity check molecular switch circuit is shown below. Figure 5 As shown, the DNA chemical reaction rate was set to 1.0E-4 nMs throughout the entire reaction process. -1 The leakage rate of the switch in the blocked state is set to 1.0E-6nM. -1 s -1 The leakage rate of the switch in the open state is set to 5.0E-6nM. -1 s -1 The initial concentration of the molecular switches is set to 1xnM, where 1x represents 100nM. A high-concentration signal chain will only be obtained at the output when all switches in any path throughout the reaction receive a high-concentration signal chain.

[0178] Simulation results are as follows Figure 6 As shown, when the input terminal ABC is 000, the odd parity output terminal Σ O It outputs a high-concentration fluorescence signal and reaches dynamic equilibrium within 600 seconds, while the even-calibrated output terminal Σ E The output is a low-concentration fluorescence signal, indicating that the number of 1s in the input binary 000 is even. When the input terminals ABC are 001, the odd-parity output terminal Σ... O It outputs a low-concentration fluorescence signal, while the even-parity output terminal Σ E The output is a high-concentration fluorescence signal, indicating that the number of 1s in the input binary 001 is odd. When the input terminals ABC are 010, the odd-parity output terminal Σ... O It outputs a low-concentration fluorescence signal, while the even-parity output terminal Σ EThe output is a high-concentration fluorescence signal, indicating that the number of 1s in the input binary 010 is odd. When the input terminals ABC are 011, the odd-parity output terminal Σ... O It outputs a high-concentration fluorescence signal, while the even-parity output terminal Σ E The output is a low-concentration fluorescence signal, indicating that the number of 1s in the input binary 011 is even. When the input terminal ABC is 100, the odd-parity output terminal Σ... O It outputs a low-concentration fluorescence signal, while the even-parity output terminal Σ E The output is a high-concentration fluorescence signal, indicating that the number of 1s in the input binary 100 is odd. When the input terminal ABC is 101, the odd-parity output terminal Σ... O It outputs a high-concentration fluorescence signal, while the even-parity output terminal Σ E The output is a low-concentration fluorescence signal, indicating that the number of 1s in the input binary 101 is even. When the input terminal ABC is 110, the odd-parity output terminal Σ... O It outputs a high-concentration fluorescence signal, while the even-parity output terminal Σ E The output is a low-concentration fluorescence signal, indicating that the number of 1s in the input binary 110 is even. When the input terminal ABC is 111, the odd-parity output terminal Σ... O It outputs a low-concentration fluorescence signal, while the even-parity output terminal Σ E The output is a high-concentration fluorescence signal, which means that the number of 1s in the input binary 111 is odd.

[0179] The concentration settings of all signal strands in the 3-bit parity check DNA circuit are shown in the table below:

[0180]

[0181]

[0182] Secondly, the 9-bit parity check molecular switch circuit is composed of three identical 3-bit parity check molecular switch circuit modules. The first three 3-bit parity check switch circuits output parity check current signal chains Σ respectively. O1 , Σ E1 , Σ O2 , Σ E2 and Σ O3 , Σ E3 Then, these signals are passed again as input signals to the fourth 3-bit parity check module. The chemical reaction equation for the fourth parity check module is as follows:

[0183] Σ E1 +SW1→ΣC E1 +SW1(on) (34)

[0184] ΣCE1 +DS3→DS3(off)+waste1 (35)

[0185] ΣC E1 +DS4→DS4(off)+waste2 (36)

[0186] Σ E2 +DS3(off)→ΣC E2 +DS3(on) (37)

[0187] Σ o2 +DS4(off)→ΣC O2 +DS4(on) (38)

[0188] ΣC E2 +DS7→DS7(off)+waste3 (39)

[0189] ΣC E2 +DS 10 →DS 10 (off)+waste4 (40)

[0190] ΣC O2 +DS8→DS8(off)+waste5 (41)

[0191] ΣC O2 +DS9→DS9(off)+waste6 (42)

[0192] Σ O3 +DS7(off)→Σ O +DS7(on) (43)

[0193] Σ E3 +DS8(off)→Σ O +DS8(on) (44)

[0194] Σ O1 +SW2→ΣC O2 +SW2(on) (45)

[0195] ΣC O2 +DS5→DS5(off)+waste7 (46)

[0196] ΣC O2 +DS6→DS6(off)+waste8 (47)

[0197] Σ E2 +DS5(off)→ΣC E2 +DS5(on) (48)

[0198] Σ o2 +DS6(off)→ΣC O2 +DS5(on) (49)

[0199] ΣC E2 +DS8→DS8(off)+waste9 (50)

[0200] ΣC E2 +DS9→DS9(off)+waste 10 (51)

[0201] ΣC O2 +DS7→DS7(off)+wast 11 (52)

[0202] ΣC O2 +DS 10 →DS 10 (off) + waste 12 (53)

[0203] Σ O3 +DS9(off)→Σ E +DS9(on) (54)

[0204] Σ E3 +DS 10 (off)→Σ E +DS 10 (on) (55)

[0205] Σ O +Report0→Report0(on)+waste 13 (56)

[0206] Σ E +Report1→Report1(on)+waste 14 (47)

[0207] The specific process of reaction (34)-(57) is as follows: the output strand Σ from the first three 3-bit parity check DNA switch circuits is converted into a single strand. O1 , Σ E1 , Σ O2 , Σ E2 , Σ O3 , Σ E3 This serves as the input signal for each molecular switch. When the input signal Σ O1 , Σ E1 Upon arrival, the upstream switches SW1 and SW2 of the fourth parity check DNA switch circuit close, and the output current signal chain ΣC... O1 , ΣCE1 Activate downstream switches DS3-DS6 and wait for input signal Σ. O2 , Σ E2 The arrival of Σ; when Σ O2 , Σ E2 After input, the corresponding downstream switches DS3-DS6 turn to the closed state, and the output current signal chain ΣC O2 , ΣC E2 ; Current signal chain ΣC O2 , ΣC E2 Activate downstream switch DS7-DS 10 Waiting for input signal Σ O3 , Σ E3 The arrival of Σ; when Σ O3 , Σ E3 After input, the downstream switch DS7-DS 10 Switching to a closed state, the output current signal chain Σ O , Σ E The signal is transmitted to the fluorescence reporter gates Report0 and Report1. After reacting with the fluorescence reporter gates, two fluorescence signal chains, Report0(on) and Report1(on), are output as the final verification results. One of the two fluorescence chains reaches dynamic equilibrium after 1000 seconds, while the other outputs a low fluorescence signal of less than 10 nM.

[0208] Step 5: Verify the correctness of the 9-bit parity check switch circuit using the Simbiology simulation platform.

[0209] A 9-bit parity check switch circuit model was established using the Simbiology simulation platform. With the same response rate and leakage rate set as described above, its correctness in handling the parity of 9-bit binary codes was verified. Simulation results are as follows: Figure 7 As shown, when the input terminal GHIDEFABC receives the 9-bit binary code 001010111, the output terminal Σ O Output a low fluorescence signal below 10 nM, determine if the number of 1s is odd, and output Σ. E It outputs a high fluorescence signal and reaches dynamic equilibrium after 1000 seconds.

[0210] When the input terminal GHIDEFABC receives the 9-bit binary code 001111011, the output terminal Σ O It outputs a high fluorescence signal, reaches dynamic equilibrium after 1000 seconds, and the output terminal Σ E Output a low fluorescence signal below 10nM and determine that the number of 1s is even.

[0211] When the input terminal GHIDEFABC receives the 9-bit binary code 010011000, the output terminal Σ OOutput a low fluorescence signal below 10 nM, determine if the number of 1s is odd, and output Σ. E It outputs a high fluorescence signal and reaches dynamic equilibrium after 1000 seconds.

[0212] When the input terminal GHIDEFABC receives the 9-bit binary code 011100111, the output terminal Σ O It outputs a high fluorescence signal, reaches dynamic equilibrium after 1000 seconds, and the output terminal Σ E Output a low fluorescence signal below 10nM and determine that the number of 1s is even.

[0213] When the input terminal GHIDEFABC receives the 9-bit binary code 100010001, the output terminal Σ O Output a low fluorescence signal below 10 nM, determine if the number of 1s is odd, and output Σ. E It outputs a high fluorescence signal and reaches dynamic equilibrium after 1000 seconds.

[0214] When the input terminal GHIDEFABC receives the 9-bit binary code 101011011, the output terminal Σ O It outputs a high fluorescence signal, reaches dynamic equilibrium after 1000 seconds, and the output terminal Σ E Output a low fluorescence signal below 10nM and determine that the number of 1s is even.

[0215] When the input terminal GHIDEFABC receives the 9-bit binary code 110101010, the output terminal Σ O Output a low fluorescence signal below 10 nM, determine if the number of 1s is odd, and output Σ. E It outputs a high fluorescence signal and reaches dynamic equilibrium after 1000 seconds.

[0216] When the input terminal GHIDEFABC receives the 9-bit binary code 111110100, the output terminal Σ O It outputs a high fluorescence signal, reaches dynamic equilibrium after 1000 seconds, and the output terminal Σ E Output a low fluorescence signal below 10nM and determine that the number of 1s is even.

[0217] When the input terminal GHIDEFABC receives the 9-bit binary code 111111111, the output terminal Σ O Output a low fluorescence signal below 10 nM, determine if the number of 1s is odd, and output Σ. E It outputs a high fluorescence signal and reaches dynamic equilibrium after 1000 seconds.

[0218] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for constructing a 9-bit parity check molecular switch circuit, characterized in that, The steps are as follows: Step 1: Elucidate the structure and reaction mechanism of the molecular switch circuit based on DNA strand substitution technology; Step 2: Set the reaction rate and leakage rate, and optimize the input signal of the molecular switch circuit and the concentration ratio of each stage of the molecular switch; Step 3: Set up the fan-in molecular switch circuit and the fan-out molecular switch circuit according to the molecular switch circuit; Step 4: Using the switch canvas routing strategy, build a 3-bit parity check molecular switch circuit based on the fan-in molecular switch circuit and the fan-out molecular switch circuit. Use four identical 3-bit parity check molecular switch circuits to build a 9-bit parity check molecular switch circuit. Step 5: Verify the correctness of the 9-bit parity check molecular switch circuit using the Simbiology simulation platform; The molecular switch circuit in step one includes a start switch SW, a downstream switch DS that receives upstream signals, and a fluorescence reporter gate Report; the chemical reaction mechanism of the molecular switch circuit is as follows: A+SW→CA+SW(on) (1); CA+DS→DS(off)+waste1 (2); B+DS(off)→CY+DS(on) (3); CY+Report→Report(on)+ waste2 (4); Among them, reaction (1) is the chain substitution reaction between input signal A and start switch SW, where A and SW are both reactants, CA and SW(on) are both products, CA represents the generated current signal chain, and SW(on) represents the start switch SW being closed; reaction (2) is the chain substitution reaction between current signal chain CA and downstream switch DS which is in a blocked state, where CA and DS are both reactants, DS(off) and waste1 are both new products, DS(off) represents the downstream switch DS changing from the original blocked state to the open state, and waste1 is waste; reaction (3) is the chain substitution reaction between input signal B and switch DS(off), producing products CY and DS(on), where CY is the current signal chain, and DS(on) represents the downstream switch DS changing to the closed state; reaction (4) is the chain substitution reaction between current signal chain CY and fluorescence reporter gate Report, where Report(on) is the generated fluorescence signal chain, and represents the fluorescence reporter gate Report being closed. This reaction process converts the concentration of signal chain CY into fluorescence intensity output; The reaction process is as follows: Input signal A enters from the input terminal and combines with the exposed foothold of start switch SW, thereby generating a current signal chain CA; downstream switch DS consists of S-domain and D-domain. S-domain receives the current signal chain CA transmitted from upstream switch through the exposed foothold, while D-domain responds to the input signal B of downstream switch. After the current signal chain CA is replaced by the downstream switch DS chain, the downstream switch DS is switched to the open state, waiting for the arrival of input signal Y; after the downstream switch DS is replaced by the input signal Y, a current signal chain CY is generated, and the downstream switch DS is switched to the closed state DS(on). Finally, the current signal chain CY reacts with the fluorescence reporter gate to generate a fluorescence signal chain that is detected and output. The fan-in molecular switch circuit includes a 2-fan-in molecular switch circuit, which aggregates two current signals transmitted from the upstream switch into a single current signal. The 2-fan-in molecular switch circuit includes upstream switches DS1 and DS2, and downstream switch SW1. Upstream switches DS1 and DS2 are connected in parallel and then in series with the downstream switch SW1. The reaction equation for the 2-fan-in molecular switch circuit is: A1+DS1→CA1+DS1(on) (5); A2+DS2→CA1+DS2(on) (6); CA1+SW→SW (off) (7); B+SW (off)→CS+SW(on) (8); In reactions (5) and (6), the input signals A1 and A2 undergo chain substitution reactions with the upstream switches DS1 and DS2 respectively, generating the same current signal chain CA1, and switching the states of the downstream switches DS1 and DS2 to the closed states DS1(on) and DS2(on) respectively. In reaction (7), the current signal chain CA1 undergoes chain substitution reactions with the downstream switch SW, and the current signal chain CA1 activates the downstream switch SW. The state of the downstream switch SW changes from the original blocked state to the open state SW1(off), exposing the middle small fulcrum to wait for the arrival of the input signal B. In reaction (8), the input signal B undergoes chain substitution reactions with the downstream switch SW(off) to generate a current signal chain CS, and the state of the downstream switch SW changes to the closed state SW(on). The fan-out molecular switch circuit includes a 2-fan-out molecular switch circuit, which includes an upstream switch SW and two downstream switches DS1 and DS2 connected in parallel. The upstream switch SW is connected in series with the parallel downstream switches DS1 and DS2. The reaction equation for the 2-fan-out molecular switch circuit is: A+SW→CA+SW(on) (9); CA+DS1→DS1(off)+waste1 (10); CA+DS2→DS2(off)+waste2 (11); B1+DS1(off)→CB+DS1(on) (12); B2+DS2(off)→CB+DS2(on) (13); In reaction (9), the input signal A is replaced by the upstream switch SW chain to generate the current signal chain CA, and the upstream switch SW is converted to the closed state SW(on); in reaction (10)-(11), the two fan-out current signal chains CA are simultaneously transmitted to the downstream switches DS1 and DS2 and activate them, and the states of the downstream switches DS1 and DS2 are converted to the open states DS1(off) and DS2(off) respectively; in reaction (12)-(13), the input signals B1 and B2 react with the exposed DS1(off) and DS2(off) with the middle small support point respectively to generate the same current signal chain CB, double chain DS1(on) and double chain DS2(on), and the downstream switches DS1 and DS2 are converted from the open state to the closed state. At this time, the current signal chain CB is the output signal of the 2-fan-out molecular switch circuit.

2. The method for constructing a 9-bit parity check molecular switch circuit according to claim 1, characterized in that, In the reaction formula (1), the DNA single-strand structure of the input signal A is as follows:<n1^ s1^ s2^ n2^ n3^> The DNA double-stranded structure of the start switch SW is<t2^ t1 t2^> [n1^ s1^ s2^ n2^]{n3^*}, the single-stranded DNA structure of the product CA is as follows:<t2^ t1 t2^n1^ s1^ s2^ n2^> The DNA double-stranded structure of the closed switch SW(on) is [n1^ s1^ s2^ n2^ n3^]; in the reaction formula (2), the DNA double-stranded structure of the downstream switch DS is<u3^ v3 u4^> [u1^ v1^ v2^]:[u5^ t2^ t1 t2^]{n1^*}, the DNA double-stranded structure of the product DS(off) is as follows:<u3^ v3 u4^> [u1^ v1^ v2^]:{u5^}:[t2^ t1 t2^ n1^]<s1^ s2^ n2^> In the reaction formula (3), the DNA single-strand structure of the input signal Y is as follows:<u1^ v1^ v2^ u5^> After the input signal B undergoes a strand substitution reaction with DS(off), the single-stranded DNA structure of the product CY is as follows:<u3^ v3^ u4^ u1^v1^ v2^ > The DNA double-stranded structure of the product DS(on) is [u1^ v1^ v2^ u5^]:[ t2^ t1 t2^ n1^]<s1^s2^ n2^> In the reaction formula (4), the DNA double-stranded structure of the fluorescent reporter gate Report is [u1^ v3 u4^ ]{u1^*}, and the DNA double-stranded structure of the product Report(on) is [u1^ v3 u4^ u1^]{ v1^ v2^}; where s1, s2, n2, t1, t2, u3, u4, v1, v2, v3 are structural domains, u1, n1, n3, u5 are small fulcrum domains, < > represents the upper strand structure of the DNA strand, [ ] represents the double-stranded structure of the DNA strand that has been complementary, : represents the double-stranded structural domain used to connect the two double-stranded structural domains, {} represents the lower strand structure of the DNA strand, ^ is used to mark the upper strand structural domain, and ^* is used to mark the lower strand structural domain; In step two, the reaction rate of the molecular switch circuit is set to 1.0E-4nMs. -1 The leakage rate of the switch in the blocked state is set to 1.0E-6 nM. -1 s -1 The leakage rate of the switch in the open state is set to 5.0E-6 nMs. -1 The initial concentration of input signal A is set to twice the concentration of the start switch SW. If the upstream switch and the downstream switch are connected in series, the initial concentration ratio of the molecular switches is set to 1.5x:1x, where x=100nM.

3. The method for constructing a 9-bit parity check molecular switch circuit according to claim 1, characterized in that, In the reaction equations (6) and (7) of the 2-fan-in molecular switch circuit, the DNA single-strand structures of the input signals A1 and A2 are designed as follows:<n1^ s1^ s2^ n2^ n3^> and<n1^ s1^ s2^ n2^ n4^> The DNA double-stranded structures of upstream switches DS1 and DS2 are respectively<t2^ t1 t2^> [n1^ s1^ s2^ n2^]{n3^*} and<t2^ t1 t2^> [n1^ s1^ s2^ n2^]{n4^*}, the single-stranded DNA structure of the product CA1 is as follows:<t2^ t1 t2^ n1^ s1^ s2^ n2^> The DNA double-stranded structure of the downstream switch SW1 in reaction (8) is as follows:<u2^ v3 u3^> [u1^ v1^ v2^]:[u4^ t2^ t1 t2^]{n1^*};The single-stranded DNA structure of the input signal strand Y in reaction formula (9) is<u1^ v1^ v2^ u4^> The generated current signal chain CS is a single-stranded DNA.<u2^ v3u3^ u1^ v1^ v2^> ; where t1, t2, s1, s2, n2, u1, u2, u3, v1, v2, v3 are structural domains, n1, n3, n4, u4 are small fulcrum domains, < > represent the upper strand structure of the DNA strand, [ ] represent the double-stranded structure of the DNA strand that has been complementary, : represents the double-stranded structural domain used to connect the two double-stranded structural domains, {} represents the lower strand structure of the DNA strand, ^ is used to mark the upper strand structural domain, and ^* is used to mark the lower strand structural domain; The initial concentrations of upstream switches DS1 and DS2 are both set to 1.5x, the initial concentration of downstream switch SW1 is set to 1x, the initial concentrations of all input signals are set to twice the initial concentration of the molecular switch, and the initial concentration of input signal X1 of upstream switch DS1 is set to 3x. In the reaction equation (10) of the 2-fan-out molecular switch circuit, the DNA single-strand structure of the input signal A is as follows:<n1^ s1^ s2^n2^ n3^> The DNA double-stranded structure of the upstream switch SW is<t2^ t1 t2^> [n1^ s1^ s2^ n2^]{n3^*}, the single-stranded DNA structure of the product CA is as follows:<t2^ t1 t2^ n1^ s1^ s2^ n2^> The DNA double-stranded structures of downstream switch DS1 and downstream switch DS2 in the reaction formulas (11)-(12) are respectively<u2^ v3 u3^> [u1^ v1^ v2^]:[u4^ t2^ t1 t2^]{n1^*}sum<u2^ v3 u3^> [u1^ v1^ v2^]:[u5^ t2^ t1 t2^]{n1^*};The DNA single-strand structures of the input signals Y1 and Y2 in the reaction formulas (13)-(14) are respectively<u1^ v1^ v2^ u4^> and<u1^ v1^ v2^ u5^> The generated current signal chain CS is<u2^ v3 u3^ u1^ v1^ v2^> ; where s1, s2, n2, u1, u3, t1, t2, v1, v2, v3 are structural domains, n1, n3, u4, u5 are small fulcrum domains, < > represents the upper strand structure of the DNA strand, [ ] represents the double-stranded structure of the DNA strand that has been complementary, : represents the double-stranded structural domain used to connect the two double-stranded structural domains, {} represents the lower strand structure of the DNA strand, ^ is used to mark the upper strand structural domain, and ^* is used to mark the lower strand structural domain; The initial concentration of the upstream switch SW2 is set to 1.5x, the initial concentration of the downstream switches DS3 and DS4 is set to 1x, the initial concentration of all input signals is set to twice the initial concentration of the molecular switch, and the initial concentration of the input signal A corresponding to the upstream switch SW2 is set to 3x.

4. The method for constructing a 9-bit parity check molecular switch circuit according to any one of claims 1-3, characterized in that, The method for constructing the 3-bit parity check molecular switch circuit is as follows: a 3-bit parity check switch circuit is constructed using a switch canvas strategy; a DNA switch circuit with two layers of 10 molecular switches is designed based on the DNA strand substitution reaction. The 9-bit parity check molecular switch circuit includes four identical 3-bit parity check molecular switch circuits. The parity of the three identical 3-bit parity check molecular switch circuits is detected by detecting the parity of the three sets of binary codes respectively. The output current signal chain is used as the input signal of the fourth parity check module to perform the final parity check and finally obtain the check result.

5. The method for constructing a 9-bit parity check molecular switch circuit according to claim 4, characterized in that, The 3-bit parity check molecular switch circuit includes input start switches SW1 and SW2, intermediate downstream switches DS3-DS6, and output downstream switches DS7-DS6. 10 And two fluorescent reporter gates, Report0 and Report1, whose DNA strand substitution reactions include: The process of input signal A' activating start switch SW1 is as follows: A'+SW1→CSA'+SW1(on) (14); The reaction of the 2-fan-out molecular switch circuit is: CSA'+DS3→DS3(off)+waste1 (15); CSA'+DS4→DS4(off)+waste2 (16); In reaction (14), after the input signal A' is chain-replaced with the start switch SW1, a current signal chain CSA' is generated, and the start switch SW1 is turned into the closed state SW1(on); in reaction (15)-(16), the current signal chain CSA' is fanned out twice and then transmitted to the downstream switch DS3 and downstream switch DS4 which are in the blocked state. The downstream switch DS3 and downstream switch DS4 are turned into the open state DS3(off) and DS4(off) respectively, with the products waste1 and waste2 being waste materials; The process by which input signal B' and input signal B activate downstream switches DS3 and DS4 respectively and output current signal chains is as follows: B'+DS3(off)→CSB'+DS3(on) (17); B+DS4(off)→CSB+DS4(on) (18); In reaction formulas (17)-(18), input signal B' and input signal B react chemically with downstream switches DS3(off) and DS4(off) which are in the off state, respectively, generating two current signal chains CSB' and CSB, which are respectively transmitted to downstream switches DS7 and DS8. 10 At downstream switches DS8 and DS9, and change downstream switches DS3 and DS4 from open to closed states DS3(on) and DS4(on); Input signal A activates start switch SW2 as follows: A+SW2→CSA+SW2(on) (19); The reaction of the 2-fan-out molecular switch circuit is: CSA+DS5→DS5(off)+waste3 (20); CSA+DS6→DS6(off)+waste4 (21); In reaction (19), after the input signal A is chain-replaced with the start switch SW2, a current signal chain CSA is generated, and the start switch SW2 is turned into the closed state SW2(on); in reactions (20)-(21), the current signal chain CSA is fanned out twice and then transmitted to the downstream switches DS5 and DS6 which are in the blocked state. The downstream switches DS5 and DS6 are turned into the open state DS3(off) and DS4(off) with the middle exposed small fulcrum. The products waste1 and waste2 are waste materials; The process by which input signal B' and input signal B react chemically with downstream switches DS5(off) and DS6(off), which are in the off state, is as follows: B'+DS5(off)→CSB'+DS5(on) (22); B+DS6(off)→CSB+DS6(on) (23); In reaction formulas (22)-(23), input signal B' and input signal B react chemically with downstream switches DS5(off) and DS6(off) which are in the off state, respectively, to generate two current signal chains CSB' and CSB, which are transmitted to downstream switches DS8 and DS9 and downstream switches DS7 and DS8 respectively. 10 At this point, the downstream switches DS5 and DS6 are changed from the open state to the closed state DS5(on) and DS6(on); The upstream current signal chains CSB' and CSB activate the downstream switches DS7 and DS, respectively. 10 The reaction process with DS8 and DS9 is as follows: CSB'+DS7→DS7(off)+waste5 (24); CSB'+DS 10 →DS 10 (off)+waste6 (25); CSB+DS8→DS8(off)+waste7 (26); CSB+DS9→DS9(off)+waste8 (27); Among them, waste5, waste6, waste7, and waste8 are waste materials, and the activated downstream switches DS7, DS8, DS9, and DS... 10 The previously blocked states were respectively switched to open states: DS7(off), DS8(off), DS9(off), DS 10 (off), exposing the small central pivot point to await the arrival of input signals C and C'; Input signals C', C', C', and C' are respectively connected to downstream switches DS7(off), DS8(off), DS9(off), and DS'(off) which are in the off state. 10 The process by which (off) produces a chemical reaction is as follows: C'+DS7(off)→Σ O +DS7(on) (28); C+DS8(off)→Σ O +DS8(on) (29); C'+DS9(off)→Σ E +DS9(on) (30); C+DS 10 (off)→Σ E +DS 10 (on) (31); Among them, downstream switches DS7~DS 10 The switches transition from the open state to the closed state respectively: DS7(on), DS8(on), DS9(on), DS 10 (on), Σ O , Σ E For the two generated current signal chains, current signal chain Σ O , Σ E It is transmitted to the fluorescence reporting gates Report0 and Report1; Current signal chain Σ O With the fluorescence report gate Report0 and the current signal chain Σ E The reaction processes with the fluorescent reporter gate Report1 are as follows: Σ O +Report0→Report0(on)+waste9 (32); S E +Report1→Report1(on)+waste 10 (33); The closure of the two fluorescent reporter gates, Report0 and Report1, generates two fluorescent signal chains, Report0(on) and Report1(on), respectively, producing waste9 and waste. 10 Waste material; The DNA chemical reaction rate is set to 1.0E-4 nMs in the 3-bit parity check molecular switch circuit. -1 The leakage rate of the switch in the blocked state is set to 1.0E-6 nMs. -1 The leakage rate of the switch in the open state is set to 5.0E-6 nMs. -1 The initial concentration of the molecular switch is set to 1xnM, where x represents 100. The initial concentrations of input signals A, B, and C are set to 2xnM. The concentration ratio of each stage of the molecular switch is set to 4:1.5:

1.

6. The method for constructing a 9-bit parity check molecular switch circuit according to claim 5, characterized in that, The chemical reaction equation for the fourth parity check module is: S E1 + SW1→ΣC E1 + SW1(on)(34); ΣC E1 +DS3→DS3(off)+waste1(35); ΣC E1 +DS4→DS4(off)+waste2(36); Σ E2 +DS3(off)→ΣC E2 +DS3(on)(37); Σ o2 +DS4(off)→ΣC O2 +DS4(on) (38); ΣC E2 +DS7→DS7(off)+waste3(39); ΣC E2 +DS 10 →DS 10 (off)+waste4(40); ΣC O2 +DS8→DS8(off)+waste5(41); ΣC O2 +DS9→DS9(off)+waste6(42); Σ O3 + DS7(off)→CY O +DS7(on) (43); Σ E3 + DS8(off)→CY O +DS8(on) (44); S O1 + SW2→ΣC O2 + SW2(on) (45); ΣC O2 +DS5→DS5(off)+waste7(46); ΣC O2 +DS6→DS6(off)+waste8(47); S E2 +DS5(off)→ΣC E2 +DS5(on) (48); Σ o2 +DS6(off)→ΣC O2 +DS5(on) (49); ΣC E2 +DS8→DS8(off)+waste9(50); ΣC E2 +DS9→DS9(off)+waste 10 (51); ΣC O2 +DS7→DS7(off)+wast 11 (52); ΣC O2 +DS 10 →DS 10 (off)+waste 12 (53); Σ O3 + DS9(off)→CY E +DS9(on)(54); Σ E3 + DS 10 (off)→CY E +DS 10 (on) (55); S O +Report0→Report0(on)+waste 13 (56); S E + Report1→Report1(on)+waste 14 (57); Among them, the product waste1-waste 14 As waste, the reaction process of reaction (34)-(57) is as follows: the output chain Σ from the first three 3-bit parity check molecular switch circuits is converted into waste. O1 Σ E1 Σ O2 Σ E2 Σ O3 Σ E3 As the input signal chain for each molecular switch; when the output chain Σ O1 Σ E1 Upon arrival, upstream switches SW1 and SW2 close, and the output current signal chain ΣC is activated. O1 , ΣC E1 Activate downstream switches DS3-DS6, and wait for the output chain Σ. O2 Σ E2 The input; when the output chain Σ O2 Σ E2 After input, the corresponding downstream switches DS3-DS6 turn to the closed state, and the output current signal chain ΣC O2 , ΣC E2 ; Current signal chain ΣC O2 , ΣC E2 Activate downstream switch DS7-DS 10 Waiting for the output chain Σ O3 Σ E3 The input; when the output chain Σ O3 Σ E3 After the input, the downstream switch DS7-DS 10 Switching to a closed state, the output current signal chain Σ O Σ E The signal is transmitted to the fluorescence reporter gates Report0 and Report1. After reacting with the fluorescence reporter gates, two fluorescence signal chains, Report0(on) and Report1(on), are output as the final verification results. One of the two fluorescence chains reaches dynamic equilibrium after 1000 seconds, while the other outputs a low fluorescence signal of less than 10 nM.

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