JP2014018143A - Production method of nucleic acid nanowire, nucleic acid used for the method, and nucleic acid nanowire produced by the method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a production method of a nucleic acid nanowire having sufficient characteristics relating to a thin shape, length, and stiffness.SOLUTION: The production method of a nucleic acid nanowire comprises the step of hybridizing and photocrosslinking a group of single-stranded nucleic acid for producing a nucleic acid nanowire containing a base sequence having predetermined complementarity and an artificial nucleic acid base represented by formula I.

Description

  The present invention relates to a method for producing a nucleic acid nanowire, a single-stranded nucleic acid group used in the method, and a nucleic acid nanowire produced by the method.

  DNA nanowires are considered promising as nanowires used in nanotechnology, particularly as conductive nanowirings used in next-generation semiconductor circuit miniaturization technology. Nucleic acids including DNA and RNA are natural polymers, and are very advantageous in terms of control of length and thickness compared to so-called synthetic polymers such as polyethylene and polyester. In addition, DNA nanowires are also expected to be used for sensing and the like using selectivity based on base sequences.

  However, as is well known, DNA is so flexible that long genomic DNA can be compactly housed in a very small nucleus in a cell. For this reason, DNA lacks the rigidity required for nanowires, and if simply synthesized, it bends in a compact manner and does not satisfy the requirements for nanowires as they are. Therefore, various techniques have been developed for the production of DNA nanowires.

  As a method for producing DNA nanowires, it has been proposed to produce DNA nanowires by an enzymatic method, that is, a method in which short ODNs (oligodeoxyribonucleic acids) are connected by ligase, but using an enzyme reaction From the above, it is necessary to maintain the optimum conditions of the enzyme, and the rigidity of the resulting DNA nanowire is comparable to that of a normal double helix, which is quite insufficient.

  Non-Patent Document 1 discloses an attempt to construct a DNA nanowire called G-wire using a G-quadplex structure. However, in Non-Patent Document 1, since the DNA nanowire is composed of four strands to maintain rigidity, the thickness of the obtained G-wire is four times that of the double helix. Therefore, there is a disadvantage that the obtained length is short and is only about 1 μm.

  Non-Patent Document 2 discloses an attempt to construct a DNA nanowire based on the DX tile structure. However, in Non-Patent Document 2, since it has a network structure and tries to maintain rigidity by bundling a plurality of double helices, the resulting structure has an extremely wide width of about several tens of nanometers. However, the structure is extremely difficult to control and the manufacturing operation is complicated.

  Non-Patent Document 3 discloses an attempt to construct a DNA nanowire using a DNA origami technique. However, in Non-Patent Document 3, since it is intended to maintain rigidity by bundling a plurality of double helices, the resulting structure becomes very thick with a width of about several tens of nanometers, The manufacturing operation is also complicated.

  Thus, if a plurality of double helix DNA strands are bundled in order to increase the rigidity of the DNA nanowire, the fineness necessary for the DNA nanowiring is lost. In addition, if a DNA nanowire is to be lengthened, its manufacturing operation becomes complicated, requires time, and various conditions need to be finely adjusted, which makes it difficult to manufacture. Thus, there has been no method for easily and quickly producing a DNA nanowire having sufficient characteristics in terms of thinness, length, rigidity and the like.

  A photoresponsive artificial nucleotide has been developed by the inventor's research group, and a patent application has been filed (Patent Document 1).

International Publication No. WO2009 / 066447

"A new DNA nanostructure, the G-wire, imaging scanning probe microscopy", March T C, Vesenka J, Henderson E. 1995, Nucleic Acds Res, Feb 25; 23 (4); 969-700 "DNA-Template Self-Assembly of Protein and Highly Conductive Nanowire", Hao Yan, Sung Ha Park, Gleb Finkelstein, John Relf, Thomas H Labean, 2003, Science, September 26, vol301, p1882-1884 "Interconnecting Gold Islands with DNA Origami nanotubes", Baoquan Ding, hao Wu, Wei Xu, Zhao Zhao, Yan Liu, Hongbin Yu, Hao Yan, 2010, Nano Letter, Vol10, 5065-5069

  As described above, a new method for producing a DNA nanowire (nucleic acid nanowire) having sufficient characteristics in terms of thinness, length, rigidity and the like has been demanded. Accordingly, an object of the present invention is to provide a new method for producing nucleic acid nanowires having sufficient characteristics in terms of thinness, length, rigidity, and the like.

  The present inventor has conducted intensive research on a method for producing nucleic acid nanowires. As a result, a single-stranded nucleic acid group having a predetermined complementary base sequence is hybridized as a monomer, and these monomers are used. Has a vinyl carbazole structure that has been introduced beforehand at a predetermined position in the double-stranded multimer by extending a double-stranded multimer in a double helix by forming a complementary base pair. By irradiating a photoresponsive artificial base with light to form a photocrosslink, it was found that this double-stranded multimer becomes a nucleic acid nanowire, and the present invention has been achieved.

  The nucleic acid nanowire thus obtained has sufficient characteristics in terms of thinness, length, rigidity and the like, and has been sought for a long time. In addition, the photocrosslinking for producing the nucleic acid nanowire is formed by light irradiation of about several seconds if the above-described photoresponsive artificial base is introduced. Do not need. Therefore, the present invention newly provides a simple and rapid method for producing nucleic acid nanowires. Furthermore, the present invention newly provides a group of single-stranded nucleic acids used for the production of the nucleic acid nanowires.

Accordingly, the present invention also includes the following (1) to (1).
(1)
A single-stranded nucleic acid group for producing nucleic acid nanowires,

Single-stranded nucleic acid group is
The first single-stranded nucleic acid M ₁ ,
The second single-stranded nucleic acid M ₂ ,
The third single-stranded nucleic acid M ₃ ,
...
n-th single-stranded nucleic acid M _n ,
n + 1st single-stranded nucleic acid M _{n + 1} ,
Z-th single-stranded nucleic acid M _z which is the last nucleic acid
(Where n is 0 or an even number greater than or equal to 2 and smaller than z;
n + 1 is an odd number of 1 or more and smaller than z;
z is an integer of 1 or more)
Comprising

The first single-stranded nucleic acid M ₁ is
It consists of a base sequence m _1a on the 5 ′ end side and a base sequence m _2a on the 3 ′ end side,
The second single-stranded nucleic acid M ₂ is
It consists of a base sequence m _3a on the 5 ′ end side and a base sequence m _2b on the 3 ′ end side,
The third single-stranded nucleic acid M ₃ is
It consists of a base sequence m _3b on the 5 ′ end side and a base sequence m _4a on the 3 ′ end side,
...
The n-th single-stranded nucleic acid M _n is (where n is 0 or an even number of 2 or more and smaller than z)
It consists of a base sequence m _{(n + 1) a} on the 5 ′ end side and a base sequence m _nb on the 3 ′ end side,
The n + 1-th single-stranded nucleic acid M _{n + 1} is (where n + 1 is an odd number of 1 or more and smaller than z)
It consists of a base sequence m _{(n + 1) b} on the 5 ′ end side and a base sequence m _{(n + 2) a} on the 3 ′ end side,
The z-th single-stranded nucleic acid M _z that is the last nucleic acid (where z is an integer of 1 or more)
If z is an even number,
It consists of a base sequence m _{(z + 1) a} on the 5 ′ end side and a base sequence m _zb on the 3 ′ end side,
If z is odd,
It consists of a base sequence m _zb on the 5 ′ end side and a base sequence m _{(z + 1) a} on the 3 ′ end side,

The first single-stranded nucleic acid M ₁ is
The sequence from the 5 ′ end to the 3 ′ end of the base sequence m _2a is a base sequence complementary to the sequence from the 3 ′ end to the 5 ′ end of the base sequence m _2b ,
The second single-stranded nucleic acid M ₂ is
The sequence from the 5 ′ end to the 3 ′ end of the base sequence m _3a is a base sequence complementary to the sequence from the 3 ′ end to the 5 ′ end of the base sequence m _3b ,
The third single-stranded nucleic acid M ₃ is
The sequence from the 5 ′ end to the 3 ′ end of the base sequence m _4a is a base sequence complementary to the sequence from the 3 ′ end to the 5 ′ end of the base sequence m _4b ,
...
The n-th single-stranded nucleic acid M _n is
Sequences 'from end 3' 5 nucleotide sequence m _{(n + 1) a} to the terminal, the base sequence m _{(n + 1)} and the sequence to the 5 'end from the 3' end of _b, with complementary nucleotide sequences Yes,
The (n + 1) th single-stranded nucleic acid M _{n + 1} is
Sequences 'from the end 3' nucleotide sequence m _{(n + 2)} 5 of _a to end, the base sequence m _{(n + 2)} and arranged to the 5 'end from the 3' end of _b, with complementary nucleotide sequences Yes,
The z-th single-stranded nucleic acid M _z that is the last nucleic acid is
The sequence from the 5 ′ end to the 3 ′ end of the base sequence m _{(z + 1) a} is a base sequence complementary to the sequence from the 3 ′ end to the 5 ′ end of the base sequence m _1a ,

The above-mentioned set of complementary base sequences has the following formula I in at least one place in at least one base sequence in each set:

(In the formula I, Ra is a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
R1 and R2 are each independently a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
N- represents a monovalent group of N. )

Having an artificial base represented by
The above-mentioned pair of complementary base sequences has any nucleobase at the position in the base sequence of the nucleic acid where the complementary base should be located with respect to the artificial base represented by the above formula I. You may,
The pair of complementary base sequences is a base at a position adjacent to the 3 ′ end side of the position in the base sequence where the complementary base should be located with respect to the artificial base represented by the above formula I. A single-stranded nucleic acid group for producing nucleic acid nanowires having a nucleobase having a pyrimidine ring.

(2)
The first single-stranded nucleic acid M ₁ is
As the base portion of at least one nucleobase in the base sequence m _2a , an artificial base represented by the formula I is included,
The second single-stranded nucleic acid M ₂ is
As the base portion of at least one nucleobase in the base sequence m _3a , an artificial base represented by the formula I is included,
The third single-stranded nucleic acid M ₃ is
As the base portion of at least one nucleobase in the base sequence m _4a , an artificial base represented by the formula I is included,
...
The n-th single-stranded nucleic acid M _n is
As the base portion of at least one nucleobase in the base sequence m _{(n + 1) a} , an artificial base represented by the formula I
The (n + 1) th single-stranded nucleic acid M _{n + 1} is
As the base part of at least one nucleobase in the base sequence m _{(n + 2) a} , an artificial base represented by the formula I
The z-th single-stranded nucleic acid M _z that is the last nucleic acid is
The single-stranded nucleic acid group for nucleic acid nanowire production according to (1), having an artificial base represented by formula I as a base part of at least one nucleobase in base sequence m _{(z + 1) a} .

(3)
The first single-stranded nucleic acid M ₁ is
As the base portion of at least one nucleobase in the base sequence m _1a , an artificial base represented by the formula I is included,
As the base portion of at least one nucleobase in the base sequence m _2a , an artificial base represented by the formula I is included,
The third single-stranded nucleic acid M ₃ is
As the base part of at least one nucleobase in the base sequence m _3b , an artificial base represented by the formula I is included,
As the base portion of at least one nucleobase in the base sequence m _4a , an artificial base represented by the formula I is included,
...
The (n + 1) th single-stranded nucleic acid M _{n + 1} is
As the base portion of at least one nucleobase in the base sequence m _{(n + 1) b} , an artificial base represented by the formula I is included,
As the base part of at least one nucleobase in the base sequence m _{(n + 2) a} , an artificial base represented by the formula I
The z-1st single-stranded nucleic acid M _z-1 that is the nucleic acid immediately preceding the last nucleic acid is:
As the base portion of at least one nucleobase in the base sequence m _za , an artificial base represented by the formula I is included,
A single-stranded nucleic acid group for nucleic acid nanowire production, which has an artificial base represented by the formula I as a base portion of at least one nucleobase in the base sequence m _{(z-1) b} .

(4)
In the single-stranded nucleic acid group described in (1) or (2),
z is 2,
The first single-stranded nucleic acid M ₁ is a single-stranded nucleic acid A;
5 'end side nucleotide sequence m _1a is, 5' are bases sequence p _a distal,
The base sequence m _2a on the 3 ′ end side is the base sequence q _a on the 3 ′ end side,
The second single-stranded nucleic acid M ₂ is a single-stranded nucleic acid B;
The base sequence m _3a on the 5 ′ end side is the base sequence p _b on the 5 ′ end side,
The base sequence m _2b on the 3 ′ end side is the base sequence q _b on the 3 ′ end side,

A single-stranded nucleic acid A consisting of _a base sequence p _a on the 5 ′ end side and a base sequence q _a on the 3 ′ end side;
A single-stranded nucleic acid B consisting of a base sequence p _b on the 5 ′ end side and a base sequence q _b on the 3 ′ end side,

In single-stranded nucleic acid A and single-stranded nucleic acid B,
Sequences 'from end 3' 5 nucleotide sequence q _a to end, the sequence of 'from end 5' 3 base sequence q _b to the ends, a complementary nucleotide sequence,
Sequences 'from end 3' 5 nucleotide sequence p _a to the end, the sequence of 'from end 5' 3 base sequence p _b to the ends, a complementary nucleotide sequence,

Nucleic A single stranded, as the base moiety of the nucleobase of at least one location in at least the nucleotide sequence q _a, the following formula I:

(In the formula I, Ra is a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
R1 and R2 are each independently a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
N- represents a monovalent group of N. )

Having an artificial base represented by
The single-stranded nucleic acid B has any nucleobase at the position in the nucleic acid B where the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid A should be located. You may,
The single-stranded nucleic acid B is a position adjacent to the 3 ′ end side of the position in the nucleic acid B where the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid A is to be located. Having a nucleobase having a pyrimidine ring as the base of

Nucleic B of single-stranded, as at least one portion of the nucleic acid bases in at least the nucleotide sequence p _b, has an artificial base represented by the formula I,
The single-stranded nucleic acid A has any nucleobase at the position in the nucleic acid A in which the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid B should be located. You may,
The single-stranded nucleic acid A is a position adjacent to the 3 ′ end side of the position in the nucleic acid A in which the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid B is to be located. A single-stranded nucleic acid group for producing nucleic acid nanowires, having a nucleobase having a pyrimidine ring as the base.

(5)
In the single-stranded nucleic acid group described in (1) or (3),
z is 2,
The first single-stranded nucleic acid M ₁ is a single-stranded nucleic acid C;
5 'end side nucleotide sequence m _1a is, 5' are bases sequence p _a distal,
The base sequence m _2a on the 3 ′ end side is the base sequence q _a on the 3 ′ end side,
The second single-stranded nucleic acid M ₂ is a single-stranded nucleic acid D;
The base sequence m _3a on the 5 ′ end side is the base sequence p _b on the 5 ′ end side,
The base sequence m _2b on the 3 ′ end side is the base sequence q _b on the 3 ′ end side,

5 'and terminal nucleotide sequence p _a 3' and terminal nucleotide sequence q _a 1 This nucleic acid C strands consisting
A single-stranded nucleic acid D consisting of a base sequence p _b on the 5 ′ end side and a base sequence q _b on the 3 ′ end side,

In single-stranded nucleic acid C and single-stranded nucleic acid D,
Sequences 'from end 3' 5 nucleotide sequence q _a to end, the sequence of 'from end 5' 3 base sequence q _b to the ends, a complementary nucleotide sequence,
Sequences 'from end 3' 5 nucleotide sequence p _a to the end, the sequence of 'from end 5' 3 base sequence p _b to the ends, a complementary nucleotide sequence,

Nucleic acid C of single-stranded, as the base moiety of the nucleobase of at least one location in the nucleotide sequence q _a, the following formula I:

(In the formula I, Ra is a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
R1 and R2 are each independently a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
N- represents a monovalent group of N. )

Having an artificial base represented by
Nucleic acid C of single-stranded, as at least one portion of the nucleic acid bases in the nucleotide sequence p _a, having an artificial base represented by the formula I,

The single-stranded nucleic acid D has any nucleobase at the position in the nucleic acid D where the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid C should be located. You may,
The single-stranded nucleic acid D is a position adjacent to the 3 ′ end side of the position in the nucleic acid D where the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid C is to be located. A single-stranded nucleic acid group for producing nucleic acid nanowires, having a nucleobase having a pyrimidine ring as the base.

(6)
In the single-stranded nucleic acid group described in (1) or (2),
z is 1,
The first single-stranded nucleic acid M ₁ is a single-stranded nucleic acid E;
5 'end side nucleotide sequence m _1a is, 5' a base sequence r _a distal,
3 'end side nucleotide sequence m _2a is 3' to a nucleotide sequence r _b distal,

5 consists 'end of the nucleotide sequence r _a and 3' single-stranded nucleic acid E consisting of a base sequence r _b distal,

In single-stranded nucleic acid E,
Sequences 'from end 3' 5 of the base sequence r _a to end, the sequence of 'from end 5' 3 base sequence r _b to the terminal a nucleotide sequence complementary,

Single-stranded nucleic acid E as the base moiety of the nucleobase of at least one location in the nucleotide sequence r _a, the following formula I:

(In the formula I, Ra is a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
R1 and R2 are each independently a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
N- represents a monovalent group of N. )

Having an artificial base represented by
Single-stranded nucleic acid E as at least one portion of the nucleic acid bases in the base sequence r _b, has an artificial base represented by the formula I,

The single-stranded nucleic acid E has any nucleobase at the position in the nucleic acid E where the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid E should be located. You may,
The single-stranded nucleic acid E is a position adjacent to the 3 ′ end side of the position in the nucleic acid E in which the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid E is to be located. A single-stranded nucleic acid group for producing nucleic acid nanowires, having a nucleobase having a pyrimidine ring as the base.

(7)
The single-stranded nucleic acid group for producing nucleic acid nanowires according to (1), wherein z is an integer of 1 to 6.
(8)
The single-stranded nucleic acid group for nucleic acid nanowire production according to (1) to (7), wherein the length of the single-stranded nucleic acid is 12 to 120 bases.

Furthermore, this invention exists also in following (11)-.
(11)
The single-stranded nucleic acid group for producing nucleic acid nanowires according to any one of (1) to (8) is hybridized to contain a single-stranded nucleic acid contained in the single-stranded nucleic acid group as a monomer. A step of preparing a multi-stranded nucleic acid multimer,
Irradiating the prepared double-stranded nucleic acid multimer with light to form a photocrosslink between the hybridized single-stranded nucleic acid molecules to obtain nucleic acid nanowires;
A method for producing nucleic acid nanowires, comprising:
(12)
A single-stranded nucleic acid A and a single-stranded nucleic acid B contained in the single-stranded nucleic acid group for producing nucleic acid nanowires described in (4) are hybridized as monomers to form a nucleic acid A monomer. And a step of preparing a double-stranded nucleic acid AB multimer comprising a nucleic acid B monomer,
The prepared double-stranded nucleic acid AB multimer is irradiated with light to form a photocrosslink between molecules of the nucleic acid A monomer and the nucleic acid B monomer constituting the double-stranded nucleic acid AB multimer, thereby Obtaining a wire;
A method for producing nucleic acid nanowires, comprising:
(13)
A single-stranded nucleic acid C and a single-stranded nucleic acid D included in the single-stranded nucleic acid group for producing nucleic acid nanowires described in (5) are hybridized as monomers to form a nucleic acid C monomer And a step of preparing a double-stranded nucleic acid CD multimer comprising a nucleic acid D monomer,
The prepared double-stranded nucleic acid CD multimer is irradiated with light to form a photocrosslink between the nucleic acid C monomer and nucleic acid D monomer molecules constituting the double-stranded nucleic acid CD multimer, thereby Obtaining a wire;
A method for producing nucleic acid nanowires, comprising:
(14)
Double-stranded nucleic acid EE containing a nucleic acid E monomer by hybridizing single-stranded nucleic acid E contained in the single-stranded nucleic acid group for producing nucleic acid nanowires according to (6) as a monomer Preparing a multimer,
A step of obtaining a nucleic acid nanowire by irradiating the prepared double-stranded nucleic acid EE multimer with light to form a photocrosslink between molecules of the nucleic acid E monomer constituting the double-stranded nucleic acid EE multimer;
A method for producing nucleic acid nanowires, comprising:
(15)
The manufacturing method according to any one of (11) to (14), wherein the light irradiation is light irradiation including light having a wavelength of 366 nm.
(16)
The production method according to any one of (11) to (15), wherein the light irradiation is light irradiation for 0.01 to 30 seconds.
(17)
The manufacturing method in any one of (11)-(16) with which light irradiation is performed at the temperature of the range of 0-30 degreeC.
(18)
Hybridization includes a step of performing annealing by cooling at a cooling rate of 0.5 to 4 ° C./hour during cooling from at least 70 ° C. to 30 ° C., in any one of (11) to (17) Manufacturing method.
(19)
A nucleic acid nanowire produced by the production method according to any one of (11) to (18).

Furthermore, the present invention also includes the following (21) to (21).
(21)
A nucleic acid nanowire obtained by photocrosslinking a double-stranded nucleic acid multimer obtained by hybridizing the single-stranded nucleic acid group for producing a nucleic acid nanowire according to any one of (1) to (8).
(22)
The double-stranded nucleic acid AB multimer obtained by hybridizing the single-stranded nucleic acid A and the single-stranded nucleic acid B contained in the single-stranded nucleic acid group for producing nucleic acid nanowires described in (4) is photocrosslinked. Nucleic acid nanowires.
(23)
A double-stranded nucleic acid CD multimer obtained by hybridizing a single-stranded nucleic acid C and a single-stranded nucleic acid D included in the single-stranded nucleic acid group for producing nucleic acid nanowires according to (5) is photocrosslinked. Nucleic acid nanowires.
(24)
A nucleic acid nanowire obtained by photocrosslinking a double-stranded nucleic acid EE multimer hybridized with a single-stranded nucleic acid E contained in the single-stranded nucleic acid group for nucleic acid nanowire production according to (6).

  The present invention provides nucleic acid nanowires. Since the diameter of the nucleic acid nanowire according to the present invention is the diameter of the double helix of the nucleic acid, it has the smallest diameter that can be realized as a nucleic acid nanowire, and is theoretically the thinnest nanowire. Moreover, the nucleic acid nanowire according to the present invention has high rigidity and sufficient length. Thus, the present invention provides a nucleic acid nanowire having sufficient characteristics in terms of thinness, length, rigidity, etc., and provides a nucleic acid nanowire that has long been sought. .

  Moreover, according to the present invention, such excellent nucleic acid nanowires can be produced simply and rapidly. The present invention provides a practical method for producing nucleic acid nanowires that has long been desired.

FIG. 1 is a photograph of the results of denaturation PAGE of nucleic acid nanowires using two types of ODN containing ^CNV K. FIG. 2 is an AFM photograph of a nucleic acid nanowire. FIG. 3 is a height difference analysis profile of nucleic acid nanowires by AFM. FIG. 4 is an AFM photograph of many nucleic acid nanowires. FIG. 5 is a histogram showing the length distribution of nucleic acid nanowires. FIG. 6 is a modified PAGE result photograph of nucleic acid nanowires using ODN containing ^CNV K and a natural DNA sequence. FIG. 7 is a modified PAGE result photograph of nucleic acid nanowires using ODN containing ^CNV K and a natural RNA sequence. FIG. 8 is a modified PAGE photograph of nucleic acid nanowires using ODN containing ^CNV K at two locations. FIG. 9 is a histogram showing the length distribution of nucleic acid nanowires with different light irradiation times.

The present invention will be described in detail below by giving specific embodiments. The present invention is not limited to the following specific embodiments and other embodiments.
[Outline of production method of nucleic acid nanowire]
The method for producing a nucleic acid nanowire according to the present invention comprises:
A step of hybridizing a single-stranded nucleic acid group for producing nucleic acid nanowires to prepare a double-stranded nucleic acid multimer containing a single-stranded nucleic acid contained in the single-stranded nucleic acid group as a monomer,
Irradiating the prepared double-stranded nucleic acid multimer with light to form a photocrosslink between the hybridized single-stranded nucleic acid molecules to obtain nucleic acid nanowires;
A method for producing nucleic acid nanowires.

[Single-stranded nucleic acid group for nucleic acid nanowire production]
The above-mentioned single-stranded nucleic acid group for nucleic acid nanowire production is
The first single-stranded nucleic acid M ₁ ,
The second single-stranded nucleic acid M ₂ ,
The third single-stranded nucleic acid M ₃ ,
...
n-th single-stranded nucleic acid M _n ,
n + 1st single-stranded nucleic acid M _{n + 1} ,
Z-th single-stranded nucleic acid M _z which is the last nucleic acid
(Where n is 0 or an even number greater than or equal to 2 and smaller than z;
n + 1 is an odd number of 1 or more and smaller than z;
z is an integer of 1 or more)
Is included.

In the present invention, these single-stranded nucleic acids form complementary base pairs by hybridization, and are polymerized as so-called monomers to form a double helix to form a double-stranded multimer. The types of molecules of single-stranded nucleic acid used include molecular types up to the zth. The number of types of single-stranded nucleic acid molecules used, that is, z has no upper limit in principle. For example, z is 1 to 100, 1 to 50, 1 to 20, 1 to 10, 1 to 1. It can be set to an integer in the range of 8, 1-6, 1-5, 1-4, 1-3, 1-2. In order to efficiently produce nucleic acid nanopolymers with fewer kinds of materials, it is preferable that z is small. However, in order to impart a desired function and complexity, nucleic acid nanopolymers are produced using desired z. can do. The above M ₂ , M ₃ , M _n and M _{n + 1} , or n, n + 1 are parameters used for explanation, and the existence of these descriptions does not limit the range of the value of z. Absent.

  In addition to the number of types of single-stranded nucleic acid molecules to be used, which is defined by the number of z, any number of molecules of single-stranded nucleic acids having the same base sequence may be used at an arbitrary ratio. Modified molecules that have been modified can also be mixed to impart specific properties to the nucleic acid nanowires.

[Complementarity of base sequence of single-stranded nucleic acid]
The above single-stranded nucleic acid has the following base sequence:
The first single-stranded nucleic acid M ₁ is
It consists of a base sequence m _1a on the 5 ′ end side and a base sequence m _2a on the 3 ′ end side,
The second single-stranded nucleic acid M ₂ is
It consists of a base sequence m _3a on the 5 ′ end side and a base sequence m _2b on the 3 ′ end side,
The third single-stranded nucleic acid M ₃ is
It consists of a base sequence m _3b on the 5 ′ end side and a base sequence m _4a on the 3 ′ end side,
...
The n-th single-stranded nucleic acid M _n is (where n is 0 or an even number of 2 or more and smaller than z)
It consists of a base sequence m _{(n + 1) a} on the 5 ′ end side and a base sequence m _nb on the 3 ′ end side,
The n + 1-th single-stranded nucleic acid M _{n + 1} is (where n + 1 is an odd number of 1 or more and smaller than z)
It consists of a base sequence m _{(n + 1) b} on the 5 ′ end side and a base sequence m _{(n + 2) a} on the 3 ′ end side,
The z-th single-stranded nucleic acid M _z that is the last nucleic acid (where z is an integer of 1 or more)
If z is an even number,
It consists of a base sequence m _{(z + 1) a} on the 5 ′ end side and a base sequence m _zb on the 3 ′ end side,
If z is odd,
It consists of a base sequence m _zb on the 5 ′ end side and a base sequence m _{(z + 1) a} on the 3 ′ end side,
Each of these has a base sequence complementarity,
The first single-stranded nucleic acid M ₁ is
The sequence from the 5 ′ end to the 3 ′ end of the base sequence m _2a is a base sequence complementary to the sequence from the 3 ′ end to the 5 ′ end of the base sequence m _2b ,
The second single-stranded nucleic acid M ₂ is
The sequence from the 5 ′ end to the 3 ′ end of the base sequence m _3a is a base sequence complementary to the sequence from the 3 ′ end to the 5 ′ end of the base sequence m _3b ,
The third single-stranded nucleic acid M ₃ is
The sequence from the 5 ′ end to the 3 ′ end of the base sequence m _4a is a base sequence complementary to the sequence from the 3 ′ end to the 5 ′ end of the base sequence m _4b ,
...
The n-th single-stranded nucleic acid M _n is
Sequences 'from end 3' 5 nucleotide sequence m _{(n + 1) a} to the terminal, the base sequence m _{(n + 1)} and the sequence to the 5 'end from the 3' end of _b, with complementary nucleotide sequences Yes,
The (n + 1) th single-stranded nucleic acid M _{n + 1} is
Sequences 'from the end 3' nucleotide sequence m _{(n + 2)} 5 of _a to end, the base sequence m _{(n + 2)} and arranged to the 5 'end from the 3' end of _b, with complementary nucleotide sequences Yes,
The z-th single-stranded nucleic acid M _z that is the last nucleic acid is
The sequence from the 5 ′ end to the 3 ′ end of the base sequence m _{(z + 1) a} is a base sequence complementary to the sequence from the 3 ′ end to the 5 ′ end of the base sequence m _1a .
Have a complementary relationship.

By providing a base sequence having such a complementary relationship, a single-stranded nucleic acid can be hybridized on the basis of its original complementarity, so that these nucleic acids M ₁ , M ₂ , M ₃ ,. .., M _n , M _{n + 1} , and M _z form a double helix in order, and extend in a rod shape while taking a repeating structure with this group of single-stranded nucleic acids as one cycle. It forms a structure that should be called a multimer.

When z is 2 in a single-stranded nucleic acid group, for example,
Nucleic M ₁ of the first single-stranded, a nucleic acid A single-stranded consisting of the base sequence q _a 5 'nucleotide sequence p _a distal and 3' end,
The second single-stranded nucleic acid M ₂ is a single-stranded nucleic acid B composed of a base sequence p _b on the 5 ′ end side and a base sequence q _b on the 3 ′ end side,
In the single-stranded nucleic acid A and the single-stranded nucleic acid B, the sequence from the 5 ′ end to the 3 ′ end of the base sequence q _{a is} the sequence from the 3 ′ end to the 5 ′ end of the base sequence q _b . a complementary nucleotide sequence, sequence 'to end 3' 5 nucleotide sequence p _a to the end, the sequence of 'from end 5' 3 base sequence p _b to the ends, a complementary nucleotide sequence,
Have a complementary relationship.

  Thus, when z is 2, the single-stranded nucleic acids A and B form a double helix in the order of ABABAB ... by the hybridization operation based on their original complementarity, This AB is elongated in a rod shape while taking a repeating structure with one cycle, forming a structure that should be called a double-stranded AB multimer. Multimer formation by complementarity is the same even when, for example, single-stranded nucleic acids C and D are used.

In a single-stranded nucleic acid group, when z is 1, for example,
The first single-stranded nucleic acid M ₁ is a single-stranded nucleic acid E;
5 'end side nucleotide sequence m _1a is, 5' a base sequence r _a distal,
3 'end side nucleotide sequence m _2a is 3' to a nucleotide sequence r _b distal,
5 consists 'end of the nucleotide sequence r _a and 3' single-stranded nucleic acid E consisting of a base sequence r _b distal,
Nucleic Acid E single-stranded, the sequence 'to end 3' 5 of the base sequence r _a to end, the sequence of 'from end 5' 3 base sequence r _b to the terminal a nucleotide sequence complementary, Have a complementary relationship.

  Thus, when z is 1, the single-stranded nucleic acid E forms a double helix in the order of EEEEEE... By the hybridization operation based on its original complementarity. The structure is supposed to be a double-stranded EE multimer by extending in a rod shape while taking a repetitive structure with a period of 1.

  In the present invention, the length of the single-stranded nucleic acid to be used is not particularly limited as long as it is a base sequence having a length capable of forming a double-stranded multimer based on such complementarity. The length of the single-stranded nucleic acid is, for example, 12 to 120 bases, 12 to 60 bases, 14 to 50 bases, 16 to 40 bases, 16 to 32 bases, 18 to 28 bases, and 18 to 24 bases. it can. The length of each type of single-stranded nucleic acid may be the same or different as long as it satisfies the above complementary relationship. In a preferred embodiment, single stranded nucleic acids of the same length can be used. The base sequences at the 5 'end and 3' end in the single-stranded nucleic acid may be the same or different as long as they satisfy the above-mentioned complementarity relationship. In a preferred embodiment, the same length base sequence can be used.

[Photoresponsive artificial base with vinylcarbazole structure]
In the present invention, a single-stranded nucleic acid has the following formula I in order to form a photocrosslink:

Is introduced as a base portion of a nucleotide that is diester-linked in a nucleic acid, replacing the base portion of a natural nucleobase.

  Ra is a cyano group, an amide group, a carboxyl group, an alkoxycarbonyl group, or hydrogen, preferably a cyano group, an amide group, a carboxyl group, an alkoxycarbonyl group, or hydrogen, and more preferably a cyano group, an amide A group, a carboxyl group, or an alkoxycarbonyl group. The alkoxycarbonyl group is preferably C2 to C7, more preferably C2 to C6, more preferably C2 to C5, further preferably C2 to C4, further preferably C2 to C3, and particularly preferably C2. it can.

  R1 and R2 are each independently a cyano group, an amide group, a carboxyl group, an alkoxycarbonyl group, or hydrogen, preferably a cyano group, an amide group, a carboxyl group, an alkoxycarbonyl group, or hydrogen, and more preferably Is a cyano group, an amide group, a carboxyl group, or an alkoxycarbonyl group. The alkoxycarbonyl group is preferably C2 to C7, more preferably C2 to C6, more preferably C2 to C5, further preferably C2 to C4, further preferably C2 to C3, and particularly preferably C2. it can.

  N- above represents a monovalent group of N, and this part is glycosidically bonded to the 1-position of the pentasaccharide and introduced as an artificial base. That is, the following formula II:

Or a ribonucleotide represented by the following formula III:

Is introduced into a nucleic acid base sequence by a phosphodiester bond, and is represented by the following formula IV:

(Wherein Rb represents a nucleic acid strand in which a ribonucleotide or deoxyribonucleotide introduced with the group of formula I as a base moiety is introduced into a base sequence by a phosphodiester bond) Yes. As described separately, the number of artificial bases of formula I introduced into a single-stranded nucleic acid is not limited to one.

  A double helix can be formed by hybridizing a nucleic acid introduced with a photoresponsive artificial base having the above-mentioned vinylcarbazole structure and a nucleic acid having a complementary base sequence to the formed double helix. When light irradiation is performed, in the sequence of the hybridized counterpart nucleic acid, a base that is one base from the base to be paired in a complementary manner with a photoreactive artificial base; Form photocrosslinks.

  The counterpart base with which this photoresponsive artificial base can form a photocrosslink is a base having a pyrimidine ring. On the other hand, this photoresponsive artificial base does not form a photocrosslink with a base having a purine ring. That is, this photoresponsive artificial base forms a photocrosslink with cytosine, uracil, and thymine as natural nucleobases, whereas it does not form a photocrosslink with guanine and adenine. Has strong specificity. Therefore, in the present invention, the complementary relationship of each single-stranded nucleic acid satisfies this condition with respect to the photoresponsive artificial base. On the other hand, in the hybridized counterpart nucleic acid sequence, the base at the position of the base to be paired with this photoresponsive artificial base is not particularly limited as long as it does not prevent the formation of a double helix by the complementary strand. For example, any natural cytobase, uracil, thymine, guanine and adenine can be used as the natural nucleobase.

[Aspect of introduction of photoresponsive artificial base into single-stranded nucleic acid]
The set of complementary base sequences described above in the group of single-stranded nucleic acids of the present invention is an artificial base represented by the above formula I at least in one of the base sequences of each set. As the base portion of the nucleobase in the base sequence. As described above, by introducing the artificial base, all of the above-described complementary base sequence pairs can be covalently bonded to each other by photocrosslinking. In addition, for the formation of such photocrosslinking, the set of complementary base sequences described above is the base sequence of the nucleic acid where the complementary base should be located with respect to the artificial base represented by the above formula I. Any nucleobase may be present as long as it does not prevent formation of a double helix by a complementary strand, and the above-mentioned set of complementary base sequences is represented by the above formula I. It has a nucleobase having a pyrimidine ring as a base at a position adjacent to the 3 ′ end side of the position in the base sequence where a complementary base should be located with respect to the artificial base.

  In a preferred embodiment of the present invention, in the single-stranded nucleic acid group of the present invention, the photoresponsive artificial base having the above-described vinylcarbazole structure is added to all types of single-stranded nucleic acid molecules. It can be introduced as a base moiety. In another preferred embodiment, in the single-stranded nucleic acid group of the present invention, half of the molecules of all types of single-stranded nucleic acids have a photoresponse having the vinylcarbazole structure. The artificial base may be introduced as the base part of the nucleobase, and the other half may not require the introduction of the photoresponsive artificial base as the base part of the nucleobase. it can.

In a preferred embodiment, in the single-stranded nucleic acid group of the present invention, the following condition (J):
The first single-stranded nucleic acid M ₁ is
As the base portion of at least one nucleobase in the base sequence m _2a , an artificial base represented by the formula I is included,
The second single-stranded nucleic acid M ₂ is
As the base portion of at least one nucleobase in the base sequence m _3a , an artificial base represented by the formula I is included,
The third single-stranded nucleic acid M ₃ is
As the base portion of at least one nucleobase in the base sequence m _4a , an artificial base represented by the formula I is included,
...
The n-th single-stranded nucleic acid M _n is
As the base portion of at least one nucleobase in the base sequence m _{(n + 1) a} , an artificial base represented by the formula I
The (n + 1) th single-stranded nucleic acid M _{n + 1} is
As the base part of at least one nucleobase in the base sequence m _{(n + 2) a} , an artificial base represented by the formula I
The z-th single-stranded nucleic acid M _z that is the last nucleic acid is
As the base portion of at least one nucleobase in the base sequence m _{(z + 1) a} , an artificial base represented by the formula I
These single-stranded nucleic acids are complementary to the artificial base represented by the above formula I contained in the single-stranded nucleic acid having a base sequence complementary to the base sequence of each single-stranded nucleic acid. Any nucleobase may be present at a position in the base sequence of the nucleic acid to which the correct base should be located,
These single-stranded nucleic acids have a pyrimidine ring as a base at the position adjacent to the 3 ′ end of the position in the base sequence where a base complementary to the artificial base represented by the above formula I should be located. Condition (J) that the nucleic acid base has
Can be satisfied.

This condition (J), for example, when z is 2, when the case of single-stranded nucleic acid A and B described above, a nucleic acid A of one strand, at least one location in at least the nucleotide sequence q _a And the single-stranded nucleic acid B is complementary to the artificial base represented by the formula I in the single-stranded nucleic acid A. Any nucleobase may be present at the position in the nucleic acid B where a simple base should be located, and the single-stranded nucleic acid B is represented by the above formula I in the single-stranded nucleic acid A. A nucleic acid base having a pyrimidine ring as a base at the position adjacent to the 3 ′ end of the position in the nucleic acid B in which a base complementary to the artificial base to be located is located, as at least one portion of the nucleic acid bases in the nucleotide sequence p _b, has an artificial base represented by the above formula I, the single-stranded nucleic acid May have any nucleobase at the position in the nucleic acid A where the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid B should be located, The single-stranded nucleic acid A is a position adjacent to the 3 ′ end side of the position in the nucleic acid A in which the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid B is to be located. It has a nucleobase having a pyrimidine ring as the base. If such conditions are satisfied, the necessary photocrosslinking is formed, and therefore an artificial base represented by the formula I can be further introduced as desired, such as rigidity.

In a preferred embodiment, in the single-stranded nucleic acid group of the present invention, the following condition (K):
The first single-stranded nucleic acid M ₁ is
As the base portion of at least one nucleobase in the base sequence m _1a , an artificial base represented by the formula I is included,
As the base portion of at least one nucleobase in the base sequence m _2a , an artificial base represented by the formula I is included,
The third single-stranded nucleic acid M ₃ is
As the base part of at least one nucleobase in the base sequence m _3b , an artificial base represented by the formula I is included,
As the base portion of at least one nucleobase in the base sequence m _4a , an artificial base represented by the formula I is included,
...
The (n + 1) th single-stranded nucleic acid M _{n + 1} is
As the base portion of at least one nucleobase in the base sequence m _{(n + 1) b} , an artificial base represented by the formula I is included,
As the base part of at least one nucleobase in the base sequence m _{(n + 2) a} , an artificial base represented by the formula I
The z-1st single-stranded nucleic acid M _z-1 that is the nucleic acid immediately preceding the last nucleic acid is:
As the base portion of at least one nucleobase in the base sequence m _za , an artificial base represented by the formula I is included,
As the base part of at least one nucleobase in the base sequence m _{(z-1) b} , an artificial base represented by the formula I is included,
These single-stranded nucleic acids are complementary to the artificial base represented by the above formula I contained in the single-stranded nucleic acid having a base sequence complementary to the base sequence of each single-stranded nucleic acid. Any nucleobase may be present at a position in the base sequence of the nucleic acid to which the correct base should be located,
These single-stranded nucleic acids have a pyrimidine ring as a base at the position adjacent to the 3 ′ end of the position in the base sequence where a base complementary to the artificial base represented by the above formula I should be located. Condition (K) that it has a nucleobase having
(However, in the condition (K), z is an even number of 2 or more)
Can be satisfied.

This condition (K), for example, when z is 2, when the case of single-stranded nucleic acid C and D described above, a nucleic acid C of single-stranded, at least one location in the nucleotide sequence q _a as the base moiety of the nucleobase having an artificial base represented by the above formula I, the nucleic acid C of single-stranded, as the base moiety of the nucleobase of at least one location in the nucleotide sequence p _a, in the formula I The single-stranded nucleic acid D having the artificial base represented is a position in the nucleic acid D where the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid C should be located The single-stranded nucleic acid D should have a base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid C. The base in the position adjacent to the 3 ′ end side of the position in the nucleic acid D has a nucleobase having a pyrimidine ring. In this case, two artificial bases are introduced into the single-stranded nucleic acid C, and no artificial base needs to be introduced into the single-stranded nucleic acid D. If such conditions are satisfied, the necessary photocrosslinking is formed, and therefore an artificial base represented by the formula I can be further introduced as desired, such as rigidity. Therefore, an artificial base may be further introduced into the single-stranded nucleic acid D.

This condition (K), for example, when z is 1, when the case of single-stranded nucleic acid E as described above, nucleic E single-stranded, at least one location of the nucleobases in the nucleotide sequence r _a as the base moiety of having an artificial base represented by the formula I, acid E single-stranded, as at least one portion of the nucleic acid bases in the base sequence r _b, artificial base represented by the formula I And the single-stranded nucleic acid E is in any position in the nucleic acid E where the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid E should be located. The single-stranded nucleic acid E may have a nucleobase, and the single-stranded nucleic acid E in the nucleic acid E in which the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid E should be located It has a nucleobase having a pyrimidine ring as the base at the position adjacent to the 3 ′ end of the position. In this case, at least one artificial base is introduced into the single-stranded nucleic acid E, and the necessary photocrosslinking is formed. An artificial base represented by I can be further introduced.

[Mode of Manufacturing Method of Nucleic Acid Nanowire]
As described above, in the method for producing nucleic acid nanowires according to the present invention, a single-stranded nucleic acid group for producing nucleic acid nanowires is hybridized to form a single-stranded nucleic acid contained in the single-stranded nucleic acid group as a monomer. A step of preparing a double-stranded nucleic acid multimer, comprising: irradiating the prepared double-stranded nucleic acid multimer with light, forming a photocrosslink between the hybridized single-stranded nucleic acid molecules, It can manufacture by the method including the process of obtaining a wire.

  In the case of the single-stranded nucleic acids A and B described above, these steps are carried out by converting the single-stranded nucleic acid A and the single-stranded nucleic acid B included in the single-stranded nucleic acid group for nucleic acid nanowire production into a single strand. A step of preparing a double-stranded nucleic acid AB multimer comprising a nucleic acid A monomer and a nucleic acid B monomer by hybridization, and irradiating the prepared double-stranded nucleic acid AB multimer with light That is, a step of forming a nucleic acid nanowire by forming a photocrosslink between the molecules of the nucleic acid A monomer and the nucleic acid B monomer constituting the double-stranded nucleic acid AB multimer.

  Further, in the case of the single-stranded nucleic acids C and D described above, these steps include the single-stranded nucleic acid C and the single-stranded nucleic acid D included in the single-stranded nucleic acid group for producing nucleic acid nanowires. , A step of preparing a double-stranded nucleic acid CD multimer containing a nucleic acid C monomer and a nucleic acid D monomer by hybridization as a monomer, and irradiating the prepared double-stranded nucleic acid CD multimer with light Thus, a step of forming a nucleic acid nanowire by forming a photocrosslink between the molecules of the nucleic acid C monomer and the nucleic acid D monomer constituting the double-stranded nucleic acid CD multimer is obtained.

  In the case of the single-stranded nucleic acid E described above, these steps are performed by hybridizing the single-stranded nucleic acid E contained in the single-stranded nucleic acid group for producing nucleic acid nanowires as a monomer. , A step of preparing a double-stranded nucleic acid EE multimer containing a nucleic acid E monomer, and light irradiation of the prepared double-stranded nucleic acid EE multimer to form a double-stranded nucleic acid EE multimer. This is a process of forming a nucleic acid nanowire by forming a photocrosslink between molecules of a monomer.

[Hybridization]
Hybridization of a single-stranded nucleic acid group for producing nucleic acid nanowires can be performed according to known procedures and conditions used to hybridize single-stranded nucleic acids to form a sequence-specific duplex. . In a preferred embodiment, it is heated to a temperature at which single-stranded nucleic acid is sufficiently dissociated, for example, heated to a temperature in the range of 88-96 ° C, 89-95 ° C, 90-94 ° C, such as 30-600. Hold for a period of time in the range of 60 to 300 seconds, fully dissociate and then slowly cool to produce sequence specific annealing, for example, during cooling from at least 70 ° C. to 30 ° C. Or during cooling from at least 70 ° C. to 25 ° C., or during cooling from at least 70 ° C. to 4 ° C., for example, 0.5-4 ° C./hour, 0.7-2 ° C./hour, 0.8 Hybridization can be performed by annealing at a cooling rate of ˜1.2 ° C./hour and annealing. In a preferred embodiment, the cooling can be performed until the temperature at which the light irradiation is performed is reached.

[Light irradiation]
The light irradiation for forming the photocrosslink can use light generally having a wavelength in the range of 330 to 370 nm, preferably 330 to 360 nm. In a preferred embodiment, light having a wavelength of 366 nm, particularly preferably, laser light having a single wavelength of 366 nm can be used. In order to advance the photocrosslinking reaction, it is generally 0-50 ° C, preferably 0-40 ° C, more preferably 0-30 ° C, such as 0-25 ° C, 4-30 ° C, 4-25 ° C. The light irradiation can be performed at a temperature in the range. In a preferred embodiment, when DNA is used as a single-stranded nucleic acid, for example, 0 to 30 ° C., 4 to 25 ° C., 0 to 10 ° C., 0 to 6 ° C., 0 to 5 ° C., 0 to 4 Light irradiation can be performed at a temperature in the range of ° C. In a preferred embodiment, when RNA is used as a single-stranded nucleic acid, for example, 0 to 30 ° C, 4 to 30 ° C, 4 to 25 ° C, 10 to 30 ° C, 15 to 30 ° C, 20 to 30 ° C. Light irradiation can be performed at a temperature in the range of 20 ° C to 20 ° C. This photocrosslinking is advantageous in that there are no particular restrictions on pH, salt concentration, etc. during light irradiation because of the use of a photoreaction. Since the formation of this photocrosslinking proceeds very rapidly by light irradiation, the time of light irradiation is sufficiently short compared with an enzyme reaction or the like, for example, 0.01 to 30 seconds, 0 Light irradiation time such as 0.01 to 20 seconds, 0.01 to 20 seconds, 0.05 to 10 seconds, 0.1 to 10 seconds, 0.1 to 5 seconds, and 0.1 to 1 second can be set. . While the formation of this photocrosslinking occurs in a very short time, in the present invention, the length of the nucleic acid nanowire can be controlled by controlling the length of the light irradiation time. By lengthening the light irradiation time, the length of the nucleic acid nanowire can be increased. Photocrosslinking caused by light irradiation is a covalent bond and is sufficiently stable. For example, it is not cleaved even under strong denaturing conditions in which the nucleic acid duplex is completely dissociated.

[Nucleic acid nanowires]
Since the nucleic acid nanowire of the present invention has a diameter of about 2 nm and is as thin as a nucleic acid double helix, it achieves the smallest diameter that can be considered in principle as a nucleic acid nanowire. Yes. The length of the nucleic acid nanowire of the present invention can be from a length smaller than 1 μm to a length sufficiently longer than 5 μm. This length can be controlled by light irradiation. The nucleic acid nanowire of the present invention has been obtained as a rod-like (linear) structure by AFM observation, and has high rigidity. Under the same conditions, when an AFM observation of a DNA double helix is performed, it is normal that the nucleic acid nanowire of the present invention is almost rod-shaped (straight line) because it is usually bent and entangled in a string shape It is surprising that it has such a high rigidity that it can be observed as a wire. The reason why DNA nanowires have been considered promising as conductive nanowirings used in next-generation semiconductor circuit miniaturization technology is that the ring structure with π electrons is the center of the double helix. And exhibiting conductivity. In the nucleic acid nanowire of the present invention, the introduced artificial nucleobase has a vinyl carbazole structure, that is, does not impair the continuity of π-electron stacking. Therefore, the nucleic acid nanowire of the present invention is advantageous as a conductive nanowiring used in the next-generation semiconductor circuit miniaturization technology.

  Hereinafter, the present invention will be described in detail with reference to examples. The present invention is not limited to the examples illustrated below.

[Production Example 1]
[Synthesis of ODN having an artificial base of vinylcarbazole structure in the base moiety]
As a nucleic acid having an artificial base of the above formula I in the base moiety, the following formula V:

In order to produce ODN (oligodeoxyribonucleotide) having the nucleotide represented by ( ^CNV K) in the base sequence, synthesis was performed according to the following Scheme 1. The synthesis was performed according to the procedure disclosed in Patent Document 1 (International Publication No. WO2009 / 066447).

According to Scheme 1 above, carbazole to 3-Iodocarbazole (1), further 3-Cyanovinylcarbazole (2), further 3-Cyanovinylcarbazole-1'-β-deoxyriboside-3 ', 5'-di- (p-toluoyl ) ester (3), 3-Cyanovinylcarbazole-1'-β-deoxyriboside (4), 5'-O- (4,4'-dimethoxytrityl) -3-Cyanovinylcarbazole-1'-β-deoxyriboside (5) ) And (5'-O- (4,4'-dimethoxytrityl) -3-Cyanovinylcarbazole-1'-β-deoxyriboside-3'-O- (cyano ethoxy-N, N-diisopropylamino) phosphoramidite (6) Were synthesized in order. The amidite obtained in (6) was used for DNA synthesis by the amidite method using a DNA / RNA synthesizer (Applied Biosystems, ABI3400), and 3-cyanovinylcarbazole-1′-β-deoxyriboside (CNVK ) Containing various ODNs (ODN containing 3-cyanovinylcarbazole-1′-β-deoxyriboside ( ^CNV K)). After the synthesis, deprotection was performed for 8 hours at 55 ° C. using 28% aqueous ammonia. Then, it refine | purified by HPLC and confirmed that it was the target arrangement | sequence by mass spectrometry. The obtained ODN sequence is shown in Table 1 below.

[Example 1]
[Synthesis of nucleic acid nanowires (type 1)]
Using a single-stranded nucleic acid A (ODN A) and a single-stranded nucleic acid B (ODN B), a nucleic acid nanowire of the type (Type 1) created using two types of ODN containing ^CNV K is synthesized as follows: did. The flow of this synthesis is shown in Scheme 2 below.

As shown in Scheme 2, ODN A and ODN B have T or G as a base at a position to form a complementary base pair with respect to ^CNV K. ^CNV K has 3 of this base. A photocrosslink (indicated by an arrow) is formed on T which is a base adjacent to the terminal side.

ODN A 12.5 [mu] M and buffer ODN B 12.5 [mu] M containing ^CNV K containing ^{CNV K (100 mM NaCl, 10} mM MgCl 2, 20 mM Tris-acetate (pH7.0), 1 mM EDTA, 2% Triton-X) Annealing was performed inside. Annealing was first heated at 90 ° C. for 5 minutes and then decreased from 70 ° C. to 4 ° C. by 1 ° C. per hour. Thereafter, 366 nm light was irradiated at 4 ° C. for 0, 0.1, 0.2, 0.3, 0.4, 0.5, 1, 2, and 5 seconds using a UV-LED irradiator to obtain a photoreactant. The denaturation PAGE analysis result of the obtained photoreactant is shown in FIG.

[Denatured PAGE result]
FIG. 1 shows the results of denaturing PAGE (polyacrylamide electrophoresis) performed under the following conditions.
・ Sample conditions Dilute the sample after light irradiation 10 times with formamide containing 8M urea ・ Gel 8M urea, 25% formamide, 15% acrylamide ・ Migration conditions 150 V for 120 minutes
・ Dyeing SYBRgold diluted 10,000 times for 20 minutes

Each lane in FIG. 1 is as follows from the leftmost lane (lane 1) to the rightmost lane (lane 10).
Lane 1. 25 bp DNA Lader (Ladder where a band appears every 25 mer)
Lane 2. Sample without light irradiation
Lane 3. Sample with 0.1 second light irradiation
Lane 4. Sample with 0.2 second light irradiation
Lane 5. Sample with 0.3 second light irradiation
Lane 6. Sample with 0.4 second light irradiation
Lane 7. Sample with 0.5 second light irradiation
Lane 8. Sample with 1 second light irradiation
Lane 9. Sample with 2 seconds of light irradiation
Lane 10. Sample with 5 seconds of light irradiation

  From the denaturing PAGE analysis results, only the monomer raw material can be confirmed under denaturing conditions before light irradiation (light irradiation 0 seconds), but it is confirmed that the band is shifted to the high molecular weight side by light irradiation. did it. This confirmed that single-stranded DNA strands (monomers) were linked together by the formation of crosslinks by light irradiation, and a stable multimeric structure was produced even under denaturing conditions. Next, AFM measurement was performed to confirm what structure was made. The sample after light irradiation was diluted 50-fold with water or tetrahydroxyfuran (THF), dropped 5 μl onto MICA whose surface was coated with 3-Aminopropytriethoxysilane, allowed to stand for 2 minutes, washed with 1 ml of water, then nitrogen. The water was removed by spraying and measurement with an AFM (atomic force microscope, NanoScope 3D, Veeco) was performed. AFM was measured in the atmosphere. AFM measurement results are shown in FIGS.

[AFM measurement result]
FIG. 2 is a photograph showing an AFM result of a scan size of 5 μm × 5 μm. A slightly bent rod-like (wire-like) structure is confirmed in the upper left visual field obtained by dividing the 5 μm square visual field into four parts vertically and horizontally. A line segment crossing the wire structure at a substantially right angle is shown from the upper center to the lower left of the 5 μm square field of view, and arrows are attached to two portions of the line segment across the wire.

  FIG. 3 shows the result of profiling the Z axis (height difference) along this line segment. Also in FIG. 3, arrows are attached to the same portions as the arrows in FIG. From the result of FIG. 3, it is confirmed that there is a peak with a height of about 2 nm, that is, a structure with a height of about 2 nm exists just between the arrows and at the position where the wire exists. This almost coincides with the height of 2 nm of the double-helix structure of DNA-DNA (B-type double helix), so that it was confirmed that nucleic acid nanowires of the target structure were obtained.

  FIG. 4 is a photograph showing AFM results for a larger number of nucleic acid nanowires. The field of view in FIG. 4 or a 10 μm × 10 μm square, and the bar on the lower right of the field of view indicates 1 μm. Nucleic acid nanowires were confirmed to have various lengths, all of which had a rod-like structure, and were confirmed to be rigid wires. When a DNA double helix is observed under the same conditions, it is normal that almost all of the molecules in the field of view are observed as a rigid wire because it is usually bent and entangled in a string shape. It is to be done.

  FIG. 5 is a histogram obtained by obtaining the length distribution of the nucleic acid nanowires from the AFM result of FIG. The horizontal axis indicates the length of the wire, and the vertical axis indicates the number of distributions of the length. The length of the wire was from less than 1 μm to over 5 μm, and the average length under these conditions was 2.13 μm.

[Example 2]
[Synthesis of nucleic acid nanowires (type 2)]
A single-stranded nucleic acid containing ^CNV K in two places C (ODN C), by using the single-stranded DNA consisting of only natural nucleic acid bases that do not contain ^CNV K, 1 kind of ODN containing ^CNV K in two places And a nucleic acid nanowire of type (Type 2) prepared using natural DNA was synthesized as follows. The flow of this synthesis is shown in Scheme 3 below.

As shown in Scheme 3, in this case, as the base position to make complementary base pairs with respect to ^CNV K, it has a C or G, ^CNV K 3 'end of the base A photocrosslink (indicated by an arrow) is formed with respect to T which is a base adjacent to the side.

Natural DNA sequence without ^CNV K (5'-TGAGGTAGTAGTTTGTGCTGTT-3 ') 12.5 μM (corresponding to ODN D) and ODN C 12.5 μM with ^CNV K buffer (100 mM NaCl, 10 mM MgCl ₂ , 20 Annealing was performed in mM Tris-acetate (pH 7.0), 1 mM EDTA, 2% Triton-X). Annealing was first heated at 90 ° C. for 5 minutes and then decreased from 70 ° C. to 4 ° C. by 1 ° C. per hour. Thereafter, a photoreactant obtained by irradiating 366 nm light at 4 ° C. for 0, 0.1, 0.2, 0.3, 0.4, 0.5, 1, 2, 5 seconds using a UV-LED irradiator was obtained. The denaturation PAGE analysis result of the obtained photoreactant is shown in FIG.

[Denatured PAGE result]
FIG. 6 shows the results of denaturing PAGE (polyacrylamide electrophoresis) performed under the same conditions as in FIG.

Each lane in FIG. 6 is as follows from the leftmost lane (lane 1) to the rightmost lane (lane 10).
Lane 1. 25 bp DNA Lader (Ladder where a band appears every 25 mer)
Lane 2. Sample without light irradiation
Lane 3. Sample with 0.1 second light irradiation
Lane 4. Sample with 0.2 second light irradiation
Lane 5. Sample with 0.3 second light irradiation
Lane 6. Sample with 0.4 second light irradiation
Lane 7. Sample with 0.5 second light irradiation
Lane 8. Sample with 1 second light irradiation
Lane 9. Sample with 2 seconds of light irradiation
Lane 10. Sample with 5 seconds of light irradiation

From this result, it was confirmed that even DNA having a natural sequence and ODN containing ^CNV K formed a photocrosslink by light irradiation to form a multimeric structure. It was also shown that nucleic acid wires can be synthesized with natural nucleotide sequences (DNA) and ODN containing two ^CNV Ks.

[Example 3]
[Synthesis of nucleic acid nanowires (type 3)]
A single-stranded nucleic acid containing ^CNV K in two places C (ODN C), by using the single-stranded RNA consisting of only natural nucleic acid bases that do not contain ^CNV K, 1 kind of ODN containing ^CNV K in two places And a nucleic acid nanowire of type (Type 3) prepared using natural RNA was synthesized as follows. The flow of this synthesis is shown in Scheme 4 below.

As shown in Scheme 4, in this case, as the base position to make complementary base pairs with respect to ^CNV K, it has a C or G, ^CNV K 3 'end of the base A photocrosslink (indicated by an arrow) is formed with respect to U which is a base adjacent to the side.

Natural RNA sequence without ^CNV K (5'-UGAGGUAGUAGUUUGU GCUGUU-3 ') 12.5 μM and ODN C 12.5 μM with ^CNV K buffer (100 mM NaCl, 5 mM MgCl ₂ , 20 mM Tris-acetate (pH7 0.0), 1 mM EDTA, 2% Triton-X). Annealing was first heated at 90 ° C. for 5 minutes and then decreased from 70 ° C. to 25 ° C. by 1 ° C. per hour. Then, the photoreaction material which irradiated 366 nm light for 0, 0.1, 0.2, 0.3, 0.4, 0.5, 1, 2, 5, and 10 second at 25 degreeC using the UV-LED irradiation machine was obtained. The denaturation PAGE analysis result of the obtained photoreactant is shown in FIG.

[Denatured PAGE result]
FIG. 7 shows the results of denaturing PAGE (polyacrylamide electrophoresis) performed under the same conditions as in FIG.

Each lane in FIG. 7 is as follows from the leftmost lane (lane 1) to the rightmost lane (lane 11).
Lane 1. 10 bp DNA Lader (Ladder where a band appears every 10 mer)
Lane 2. Sample without light irradiation
Lane 3. Sample with 0.1 second light irradiation
Lane 4. Sample with 0.2 second light irradiation
Lane 5. Sample with 0.3 second light irradiation
Lane 6. Sample with 0.4 second light irradiation
Lane 7. Sample with 0.5 second light irradiation
Lane 8. Sample with 1 second light irradiation
Lane 9. Sample with 2 seconds of light irradiation
Lane 10. Sample with 5 seconds of light irradiation
Lane 11. Sample with 10 seconds light irradiation

From these results, it was confirmed that RNA having a natural sequence and ODN containing ^CNV K also formed a multimeric structure by forming a photocrosslink by light irradiation. It was also shown that nucleic acid wires can be synthesized with natural nucleotide sequences (RNA) and ODN containing two ^CNV Ks. In addition, the RNA used in the experiment is a natural sequence of about 20 mer, and is about the length of miRNA in vivo. Therefore, the present invention can be used for the detection of SNPs using nucleic acid nanowires. It has been shown.

[Example 4]
[Synthesis of nucleic acid nanowires (type 4)]
^{Using a} single-stranded nucleic acid E (ODN E) containing ^CNV K in two locations and using only one type of ODN containing ^CNV K in two locations, type 4 nucleic acid nanowires are Synthesized. The flow of this synthesis is shown in Scheme 5 below.

As shown in Scheme 5, in this case, as the base position to make complementary base pairs with respect to ^CNV K, we have A or T, ^CNV K 3 'end of the base A photocrosslink (indicated by an arrow) is formed with respect to T which is a base adjacent to the side.

ODN E (5'-CTTCCGGA ^CNV KGCATGTACA ^CNV KG -3 ') containing 25 μM ^CNV K in buffer (100 mM NaCl, 10 mM MgCl ₂ , 20 mM Tris-acetate (pH7.0), 1 mM EDTA) , 2% Triton-X). Annealing was first heated at 90 ° C. for 5 minutes and then decreased from 70 ° C. to 4 ° C. by 1 ° C. per hour. Then, 366 nm light was irradiated at 4 ° C for 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1 second using a UV-LED irradiator. The denaturation PAGE analysis of the obtained light irradiation sample was performed. The denaturing PAGE analysis results are shown in FIG.

[Denatured PAGE result]
FIG. 8 shows the results of denaturing PAGE (polyacrylamide electrophoresis) performed under the same conditions as in FIG.

Each lane in FIG. 8 is as follows from the leftmost lane (lane 1) to the rightmost lane (lane 12).
Lane 1. 10 bp DNA Lader (Ladder where a band appears every 10 mer)
Lane 2. Sample without light irradiation
Lane 3. Sample with 0.1 second light irradiation
Lane 4. Sample with 0.2 second light irradiation
Lane 5. Sample with 0.3 second light irradiation
Lane 6. Sample with 0.4 second light irradiation
Lane 7. Sample with 0.5 second light irradiation
Lane 8. Sample of light irradiation 0.6
Lane 9. Light irradiation 0.7 sample
Lane 10. Light irradiation 0.8 sample
Lane 11. Sample with 0.9 second light irradiation time
Lane 12. Sample with 1 second light irradiation

From this result, it was confirmed that even when only one type of ODN containing ^CNV K in two places was used, photocrosslinking was formed by light irradiation to form a multimeric structure. It was also shown that nucleic acid wires can be synthesized even when only one type of ODN containing ^CNV K at two locations is used.

[Example 5]
[Control of length of nucleic acid nanowire by light irradiation time]
In the synthesis of nucleic acid nanowires of the above Example 1 (Type 1), AFM measurement was further performed, and a histogram similar to that in FIG. 5 was created for a sample with a different light irradiation time. The result is shown in FIG.

  The histogram of FIG. 9 shows, in order from the left end, nucleic acid nanowires manufactured with a light irradiation time of 0.5 seconds (left end), 1 second (center), and 5 seconds (right end) in the same manner as FIG. It is the histogram which measured and showed distribution number (vertical axis) for every length (horizontal axis). The average lengths were 0.73 μm (0.5 seconds), 2.08 μm (1 second), and 2.13 μm (5 seconds), respectively. As shown in FIG. 9, by increasing the light irradiation time, the length distribution of the nucleic acid nanowires moved so as to increase the length. Thus, it became clear that the length of the obtained nucleic acid nanowire can be controlled by the light irradiation time.

  ADVANTAGE OF THE INVENTION According to this invention, the nucleic acid nanowire provided with sufficient characteristics, such as thinness, length, and rigidity, can be obtained simply and in a short time. The present invention is an industrially useful invention.

Claims (13)

A single-stranded nucleic acid group for producing nucleic acid nanowires,

Single-stranded nucleic acid group is
The first single-stranded nucleic acid M ₁ ,
The second single-stranded nucleic acid M ₂ ,
The third single-stranded nucleic acid M ₃ ,
...
n-th single-stranded nucleic acid M _n ,
n + 1st single-stranded nucleic acid M _{n + 1} ,
Z-th single-stranded nucleic acid M _z which is the last nucleic acid
(Where n is 0 or an even number greater than or equal to 2 and smaller than z;
n + 1 is an odd number of 1 or more and smaller than z;
z is an integer of 1 or more)
Comprising

The first single-stranded nucleic acid M ₁ is
It consists of a base sequence m _1a on the 5 ′ end side and a base sequence m _2a on the 3 ′ end side,
The second single-stranded nucleic acid M ₂ is
It consists of a base sequence m _3a on the 5 ′ end side and a base sequence m _2b on the 3 ′ end side,
The third single-stranded nucleic acid M ₃ is
It consists of a base sequence m _3b on the 5 ′ end side and a base sequence m _4a on the 3 ′ end side,
...
The n-th single-stranded nucleic acid M _n is (where n is 0 or an even number of 2 or more and smaller than z)
It consists of a base sequence m _{(n + 1) a} on the 5 ′ end side and a base sequence m _nb on the 3 ′ end side,
The n + 1-th single-stranded nucleic acid M _{n + 1} is (where n + 1 is an odd number of 1 or more and smaller than z)
It consists of a base sequence m _{(n + 1) b} on the 5 ′ end side and a base sequence m _{(n + 2) a} on the 3 ′ end side,
The z-th single-stranded nucleic acid M _z that is the last nucleic acid (where z is an integer of 1 or more)
If z is an even number,
It consists of a base sequence m _{(z + 1) a} on the 5 ′ end side and a base sequence m _zb on the 3 ′ end side,
If z is odd,
It consists of a base sequence m _zb on the 5 ′ end side and a base sequence m _{(z + 1) a} on the 3 ′ end side,

The first single-stranded nucleic acid M ₁ is
The sequence from the 5 ′ end to the 3 ′ end of the base sequence m _2a is a base sequence complementary to the sequence from the 3 ′ end to the 5 ′ end of the base sequence m _2b ,
The second single-stranded nucleic acid M ₂ is
The sequence from the 5 ′ end to the 3 ′ end of the base sequence m _3a is a base sequence complementary to the sequence from the 3 ′ end to the 5 ′ end of the base sequence m _3b ,
The third single-stranded nucleic acid M ₃ is
The sequence from the 5 ′ end to the 3 ′ end of the base sequence m _4a is a base sequence complementary to the sequence from the 3 ′ end to the 5 ′ end of the base sequence m _4b ,
...
The n-th single-stranded nucleic acid M _n is
Sequences 'from end 3' 5 nucleotide sequence m _{(n + 1) a} to the terminal, the base sequence m _{(n + 1)} and the sequence to the 5 'end from the 3' end of _b, with complementary nucleotide sequences Yes,
The (n + 1) th single-stranded nucleic acid M _{n + 1} is
Sequences 'from the end 3' nucleotide sequence m _{(n + 2)} 5 of _a to end, the base sequence m _{(n + 2)} and arranged to the 5 'end from the 3' end of _b, with complementary nucleotide sequences Yes,
The z-th single-stranded nucleic acid M _z that is the last nucleic acid is
The sequence from the 5 ′ end to the 3 ′ end of the base sequence m _{(z + 1) a} is a base sequence complementary to the sequence from the 3 ′ end to the 5 ′ end of the base sequence m _1a ,

The above-mentioned set of complementary base sequences has the following formula I in at least one place in at least one base sequence in each set:

(In the formula I, Ra is a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
R1 and R2 are each independently a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
N- represents a monovalent group of N. )

Having an artificial base represented by
The above-mentioned pair of complementary base sequences has any nucleobase at the position in the base sequence of the nucleic acid where the complementary base should be located with respect to the artificial base represented by the above formula I. You may,
The pair of complementary base sequences is a base at a position adjacent to the 3 ′ end side of the position in the base sequence where the complementary base should be located with respect to the artificial base represented by the above formula I. A single-stranded nucleic acid group for producing nucleic acid nanowires having a nucleobase having a pyrimidine ring. In the single-stranded nucleic acid group described in claim 1,
z is 2,
The first single-stranded nucleic acid M ₁ is a single-stranded nucleic acid A;
5 'end side nucleotide sequence m _1a is, 5' are bases sequence p _a distal,
The base sequence m _2a on the 3 ′ end side is the base sequence q _a on the 3 ′ end side,
The second single-stranded nucleic acid M ₂ is a single-stranded nucleic acid B;
The base sequence m _3a on the 5 ′ end side is the base sequence p _b on the 5 ′ end side,
The base sequence m _2b on the 3 ′ end side is the base sequence q _b on the 3 ′ end side,

A single-stranded nucleic acid A consisting of _a base sequence p _a on the 5 ′ end side and a base sequence q _a on the 3 ′ end side;
A single-stranded nucleic acid B consisting of a base sequence p _b on the 5 ′ end side and a base sequence q _b on the 3 ′ end side,

In single-stranded nucleic acid A and single-stranded nucleic acid B,
Sequences 'from end 3' 5 nucleotide sequence q _a to end, the sequence of 'from end 5' 3 base sequence q _b to the ends, a complementary nucleotide sequence,
Sequences 'from end 3' 5 nucleotide sequence p _a to the end, the sequence of 'from end 5' 3 base sequence p _b to the ends, a complementary nucleotide sequence,

Nucleic A single stranded, as the base moiety of the nucleobase of at least one location in at least the nucleotide sequence q _a, the following formula I:

(In the formula I, Ra is a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
R1 and R2 are each independently a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
N- represents a monovalent group of N. )

Having an artificial base represented by
The single-stranded nucleic acid B has any nucleobase at the position in the nucleic acid B where the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid A should be located. You may,
The single-stranded nucleic acid B is a position adjacent to the 3 ′ end side of the position in the nucleic acid B where the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid A is to be located. Having a nucleobase having a pyrimidine ring as the base of

Nucleic B of single-stranded, as at least one portion of the nucleic acid bases in at least the nucleotide sequence p _b, has an artificial base represented by the formula I,
The single-stranded nucleic acid A has any nucleobase at the position in the nucleic acid A in which the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid B should be located. You may,
The single-stranded nucleic acid A is a position adjacent to the 3 ′ end side of the position in the nucleic acid A in which the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid B is to be located. A single-stranded nucleic acid group for producing nucleic acid nanowires, having a nucleobase having a pyrimidine ring as the base. In the single-stranded nucleic acid group described in claim 1,
z is 2,
The first single-stranded nucleic acid M ₁ is a single-stranded nucleic acid C;
5 'end side nucleotide sequence m _1a is, 5' are bases sequence p _a distal,
The base sequence m _2a on the 3 ′ end side is the base sequence q _a on the 3 ′ end side,
The second single-stranded nucleic acid M ₂ is a single-stranded nucleic acid D;
The base sequence m _3a on the 5 ′ end side is the base sequence p _b on the 5 ′ end side,
The base sequence m _2b on the 3 ′ end side is the base sequence q _b on the 3 ′ end side,

5 'and terminal nucleotide sequence p _a 3' and terminal nucleotide sequence q _a 1 This nucleic acid C strands consisting
A single-stranded nucleic acid D consisting of a base sequence p _b on the 5 ′ end side and a base sequence q _b on the 3 ′ end side,

In single-stranded nucleic acid C and single-stranded nucleic acid D,
Sequences 'from end 3' 5 nucleotide sequence q _a to end, the sequence of 'from end 5' 3 base sequence q _b to the ends, a complementary nucleotide sequence,
Sequences 'from end 3' 5 nucleotide sequence p _a to the end, the sequence of 'from end 5' 3 base sequence p _b to the ends, a complementary nucleotide sequence,

Nucleic acid C of single-stranded, as the base moiety of the nucleobase of at least one location in the nucleotide sequence q _a, the following formula I:

(In the formula I, Ra is a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
R1 and R2 are each independently a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
N- represents a monovalent group of N. )

Having an artificial base represented by
Nucleic acid C of single-stranded, as at least one portion of the nucleic acid bases in the nucleotide sequence p _a, having an artificial base represented by the formula I,

The single-stranded nucleic acid D has any nucleobase at the position in the nucleic acid D where the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid C should be located. You may,
The single-stranded nucleic acid D is a position adjacent to the 3 ′ end side of the position in the nucleic acid D where the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid C is to be located. A single-stranded nucleic acid group for producing nucleic acid nanowires, having a nucleobase having a pyrimidine ring as the base. In the single-stranded nucleic acid group described in claim 1,
z is 1,
The first single-stranded nucleic acid M ₁ is a single-stranded nucleic acid E;
5 'end side nucleotide sequence m _1a is, 5' a base sequence r _a distal,
3 'end side nucleotide sequence m _2a is 3' to a nucleotide sequence r _b distal,

5 consists 'end of the nucleotide sequence r _a and 3' single-stranded nucleic acid E consisting of a base sequence r _b distal,

In single-stranded nucleic acid E,
Sequences 'from end 3' 5 of the base sequence r _a to end, the sequence of 'from end 5' 3 base sequence r _b to the terminal a nucleotide sequence complementary,

Single-stranded nucleic acid E as the base moiety of the nucleobase of at least one location in the nucleotide sequence r _a, the following formula I:

(In the formula I, Ra is a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
R1 and R2 are each independently a cyano group, an amide group, a carboxyl group, a C2-C7 alkoxycarbonyl group, or hydrogen;
N- represents a monovalent group of N. )

Having an artificial base represented by
Single-stranded nucleic acid E as at least one portion of the nucleic acid bases in the base sequence r _b, has an artificial base represented by the formula I,

The single-stranded nucleic acid E has any nucleobase at the position in the nucleic acid E where the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid E should be located. You may,
The single-stranded nucleic acid E is a position adjacent to the 3 ′ end side of the position in the nucleic acid E in which the base complementary to the artificial base represented by the above formula I in the single-stranded nucleic acid E is to be located. A single-stranded nucleic acid group for producing nucleic acid nanowires, having a nucleobase having a pyrimidine ring as the base.   The single-stranded nucleic acid group for nucleic acid nanowire production according to claim 1, wherein z is an integer of 1 to 6.   The single-stranded nucleic acid group for producing nucleic acid nanowires according to any one of claims 1 to 5, wherein the single-stranded nucleic acid has a length of 12 to 120 bases. A single-stranded nucleic acid group for producing nucleic acid nanowires according to any one of claims 1 to 6, which is hybridized to contain a single-stranded nucleic acid contained in the single-stranded nucleic acid group as a monomer. Preparing a nucleic acid multimer;
Irradiating the prepared double-stranded nucleic acid multimer with light to form a photocrosslink between the hybridized single-stranded nucleic acid molecules to obtain nucleic acid nanowires;
A method for producing nucleic acid nanowires, comprising:   The manufacturing method according to claim 7, wherein the light irradiation is light irradiation including a wavelength of 366 nm.   The production method according to claim 7, wherein the light irradiation is light irradiation for 0.01 to 30 seconds.   The manufacturing method in any one of Claims 7-9 with which light irradiation is performed at the temperature of the range of 0-30 degreeC.   The production according to any one of claims 7 to 10, wherein the hybridization includes a step of annealing at a cooling rate of 0.5 to 4 ° C / hour during cooling from at least 70 ° C to 30 ° C. Method.   The nucleic acid nanowire manufactured by the manufacturing method in any one of Claims 7-11.   A nucleic acid nanowire obtained by photocrosslinking a double-stranded nucleic acid multimer hybridized with the single-stranded nucleic acid group for nucleic acid nanowire production according to any one of claims 1 to 6.