Dynamic DNA hydrogel and preparation method thereof

Abstract

The invention relates to the technical field of materials, in particular to dynamic DNA hydrogel and a preparation method thereof. According to the invention, a permanent cross-linking agent and a dynamic cross-linking agent DNA are polymerized in a photo-initiation manner to finally form the hydrogel with a three-dimensional network structure. RNA is used for attacking dynamic DNA cross-linking points, and RNsse H enzymatic reaction is used for realizing the dynamic process of the gel. The dynamic process of the gel can be accurately controlled through the action forms and the dosage of RNsse H and RNA, and dynamic control over the strength of the hydrogel is achieved. When the gel dynamically changes, the internal pore diameter of the gel also changes, so that the material can be loaded and released. DNA, RNA and RNase H have good biocompatibility, and can be used as a medical dressing; the process for preparing the hydrogel is simple and convenient, and the hydrogel can be applied to the fields of medical dressings, drug sustained release, cell-free protein synthesis and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of materials, and particularly to a dynamic DNA hydrogel and a preparation method thereof. Background Art

[0002] Non-equilibrium supramolecular assembly widely exists in biological life systems, endowing various living organisms with complex structures and special functions. Constructing a non-equilibrium supramolecular assembly system in an artificial system is not only beneficial to the understanding of life systems, but also helpful for the design and development of new soft matter materials. Among them, hydrogels are hydrophilic polymer compounds with a three-dimensional network structure. Their three-dimensional network structure is similar to biological tissues, and hydrogels have a high water content and a large drug-loading space, showing great application potential in the fields of biomedicine and drug delivery. With the in-depth research in the field of hydrogels, more precise synthesis, more sensitive stimulus responsiveness, and richer functionality have promoted researchers to develop new types of applied hydrogels. Therefore, the construction and research of artificially synthesized non-equilibrium hydrogels are of great significance.

[0003] DNA is the core genetic material of life systems, guiding the normal operation of biological development and life functions. From the perspective of material chemistry, as a natural biopolymer, DNA has characteristics that cannot be compared with synthetic polymers. Base complementarity enables DNA to have precise, diverse, and efficient self-assembly capabilities; the DNA sequence can be adjusted, enabling precise control of the DNA spatial and chemical structures, and having rich stimulus responsiveness; natural evolution has endowed organisms with a rich variety of biological enzymes that can precisely operate on DNA at the molecular level; DNA has good biocompatibility and biodegradability. Therefore, in recent years, DNA components have become the building blocks of functional polymer materials. DNA hydrogels utilize both the framework structure of hydrogels and retain the biological functions of DNA, enabling the perfect integration of the structure and function of hydrogel materials, and showing broad application prospects in the fields of biosensors, drug delivery, cell culture, protein synthesis, intelligent devices, environmental protection, etc. Summary of the Invention

[0004] In view of this, the technical problem to be solved by the present invention is to provide a dynamic DNA hydrogel and a preparation method thereof. In the present invention, a polymer is used as the backbone of the hydrogel, the chemical crosslinking between monomers is a permanent crosslinking point, and DNA is a dynamic crosslinking point, and curing is carried out by irradiating with ultraviolet light in the presence of a photoinitiator. Thereafter, the gel and RNaseH enzyme are placed in a solution, heated to open the DNA double strand, the RNase H enzyme diffuses into the pores inside the gel, and after cooling, the enzyme is fixed inside the gel to act. When needed, the hydrogel embedding the enzyme is placed in a solution, RNA is added to drive the gel, and the gel volume increases from small to large and then returns to the original state. That is: the gel is driven by RNA attacking DNA, and the gel is restored by an enzymatic reaction. During the construction of the gel, a therapeutic agent can be embedded into the gel together with the RnaseH enzyme, or the sequence of RNA or DNA can be designed to be a therapeutically active sequence, so that it itself serves as a therapeutic agent.

[0005] The dynamic DNA hydrogel provided by the present invention has the following raw materials for preparation: water-soluble monomers, DNA single strand L1 and DNA single strand L2.

[0006] The monomer serving as the backbone in the hydrogel of the present invention can be any monomer capable of forming a gel, preferably a water-soluble monomer. In some embodiments, the water-soluble monomer is an acrylic acid-based water-soluble monomer, preferably acrylamide, acrylic acid, 2-acrylamido-2-methyl-1-propanesulfonic acid, 2-hydroxyethyl acrylate acryloylglycinamide, or methacrylic acid, or at least one of them.

[0007] In the present invention, the ends of DNA single strand L1 and DNA single strand L2 both carry polymerizable monomers, so that they can be used as part of the gel backbone. In the present invention, the monomer is modified at the 3'-end or / and 5'-end of the DNA single strand. The polymerizable monomers on the two strands can both be modified at the 3'-end or both at the 5'-end, and the present invention does not limit this. Preferably, the modified polymerizable monomer is acrylamide.

[0008] In the present invention, the sequences of DNA single strand L1 and DNA single strand L2 are not limited, and any sequence that can make L1 strand and L2 strand partially complementary and does not form a hairpin structure or stem-loop structure by itself is a feasible solution. As a dynamic crosslinking point, in the present invention, the two DNA strands need to be complementary. In order to achieve dynamic crosslinking, that is, hybridization occurs under some conditions to enhance crosslinking, and hybridization does not occur under some conditions to weaken crosslinking. In the present invention, the DNA single strand L1 and DNA single strand L2 can be of equal length or unequal length, but must be partially complementary, that is, there are non-complementary segments.

[0009] In some embodiments, the length of the single-stranded DNA L1 is 10 to 100 bp, preferably 20 to 90 bp, more preferably 20 to 50 bp or 50 to 90 bp, or the length of the single-stranded DNA L1 is 20 to 40 bp, 40 to 60 bp, 60 to 80 bp; or the length of the single-stranded DNA L1 is 25 to 35 bp, 35 to 45 bp, 45 to 55 bp, 55 to 65 bp, 65 to 75 bp. In a specific embodiment, the length of the single-stranded DNA L1 is 25 bp, 26 bp, 27 bp, 28 bp, 29 bp, 30 bp, 31 bp, 32 bp, 33 bp, 34 bp or 35 bp.

[0010] In some embodiments, the length of the single-stranded DNA L2 is 10 to 100 bp, preferably 20 to 90 bp, more preferably 20 to 50 bp or 50 to 90 bp, or the length of the single-stranded DNA L2 is 20 to 40 bp, 40 to 60 bp, 60 to 80 bp; or the length of the single-stranded DNA L2 is 25 to 35 bp, 35 to 45 bp, 45 to 55 bp, 55 to 65 bp, 65 to 75 bp. In a specific embodiment, the length of the single-stranded DNA L2 is 25 bp, 26 bp, 27 bp, 28 bp, 29 bp, 30 bp, 31 bp, 32 bp, 33 bp, 34 bp or 35 bp.

[0011] In some specific embodiments, the length of DNA single strand L1 is 30 bp; the length of DNA single strand L2 is 30 bp. In some other specific embodiments, the length of DNA single strand L1 is 30 bp; the length of DNA single strand L2 is 25 bp. In some other specific embodiments, the length of DNA single strand L1 is 30 bp; the length of DNA single strand L2 is 26 bp. In some other specific embodiments, the length of DNA single strand L1 is 30 bp; the length of DNA single strand L2 is 27 bp. In some other specific embodiments, the length of DNA single strand L1 is 30 bp; the length of DNA single strand L2 is 28 bp. In some other specific embodiments, the length of DNA single strand L1 is 30 bp; the length of DNA single strand L2 is 29 bp. In some other specific embodiments, the length of DNA single strand L1 is 30 bp; the length of DNA single strand L2 is 30 bp. In some other specific embodiments, the length of DNA single strand L1 is 30 bp; the length of DNA single strand L2 is 31 bp. In some other specific embodiments, the length of DNA single strand L1 is 30 bp; the length of DNA single strand L2 is 32 bp. In some other specific embodiments, the length of DNA single strand L1 is 30 bp; the length of DNA single strand L2 is 33 bp. In some other specific embodiments, the length of DNA single strand L1 is 30 bp; the length of DNA single strand L2 is 34 bp. In some other specific embodiments, the length of DNA single strand L1 is 30 bp; the length of DNA single strand L2 is 35 bp.

[0012] In some specific embodiments, the length of the single-stranded DNA L1 is 25 bp; the length of the single-stranded DNA L2 is 30 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 25 bp; the length of the single-stranded DNA L2 is 25 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 25 bp; the length of the single-stranded DNA L2 is 26 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 25 bp; the length of the single-stranded DNA L2 is 27 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 25 bp; the length of the single-stranded DNA L2 is 28 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 25 bp; the length of the single-stranded DNA L2 is 29 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 25 bp; the length of the single-stranded DNA L2 is 30 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 25 bp; the length of the single-stranded DNA L2 is 31 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 25 bp; the length of the single-stranded DNA L2 is 32 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 25 bp; the length of the single-stranded DNA L2 is 33 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 25 bp; the length of the single-stranded DNA L2 is 34 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 25 bp; the length of the single-stranded DNA L2 is 35 bp.

[0013] In some specific embodiments, the length of the single-stranded DNA L1 is 26 bp; the length of the single-stranded DNA L2 is 30 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 26 bp; the length of the single-stranded DNA L2 is 25 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 26 bp; the length of the single-stranded DNA L2 is 26 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 26 bp; the length of the single-stranded DNA L2 is 27 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 26 bp; the length of the single-stranded DNA L2 is 28 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 26 bp; the length of the single-stranded DNA L2 is 29 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 26 bp; the length of the single-stranded DNA L2 is 30 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 26 bp; the length of the single-stranded DNA L2 is 31 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 26 bp; the length of the single-stranded DNA L2 is 32 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 26 bp; the length of the single-stranded DNA L2 is 33 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 26 bp; the length of the single-stranded DNA L2 is 34 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 26 bp; the length of the single-stranded DNA L2 is 35 bp.

[0014] In some specific embodiments, the length of the single-stranded DNA L1 is 27 bp; the length of the single-stranded DNA L2 is 30 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 27 bp; the length of the single-stranded DNA L2 is 25 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 27 bp; the length of the single-stranded DNA L2 is 26 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 27 bp; the length of the single-stranded DNA L2 is 27 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 27 bp; the length of the single-stranded DNA L2 is 28 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 27 bp; the length of the single-stranded DNA L2 is 29 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 27 bp; the length of the single-stranded DNA L2 is 30 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 27 bp; the length of the single-stranded DNA L2 is 31 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 27 bp; the length of the single-stranded DNA L2 is 32 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 27 bp; the length of the single-stranded DNA L2 is 33 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 27 bp; the length of the single-stranded DNA L2 is 34 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 27 bp; the length of the single-stranded DNA L2 is 35 bp.

[0015] In some specific embodiments, the length of DNA single strand L1 is 28 bp; the length of DNA single strand L2 is 30 bp. In some other specific embodiments, the length of DNA single strand L1 is 28 bp; the length of DNA single strand L2 is 25 bp. In some other specific embodiments, the length of DNA single strand L1 is 28 bp; the length of DNA single strand L2 is 26 bp. In some other specific embodiments, the length of DNA single strand L1 is 28 bp; the length of DNA single strand L2 is 27 bp. In some other specific embodiments, the length of DNA single strand L1 is 28 bp; the length of DNA single strand L2 is 28 bp. In some other specific embodiments, the length of DNA single strand L1 is 28 bp; the length of DNA single strand L2 is 29 bp. In some other specific embodiments, the length of DNA single strand L1 is 28 bp; the length of DNA single strand L2 is 30 bp. In some other specific embodiments, the length of DNA single strand L1 is 28 bp; the length of DNA single strand L2 is 31 bp. In some other specific embodiments, the length of DNA single strand L1 is 28 bp; the length of DNA single strand L2 is 32 bp. In some other specific embodiments, the length of DNA single strand L1 is 28 bp; the length of DNA single strand L2 is 33 bp. In some other specific embodiments, the length of DNA single strand L1 is 28 bp; the length of DNA single strand L2 is 34 bp. In some other specific embodiments, the length of DNA single strand L1 is 28 bp; the length of DNA single strand L2 is 35 bp.

[0016] In some specific embodiments, the length of DNA single strand L1 is 29 bp; the length of DNA single strand L2 is 30 bp. In some other specific embodiments, the length of DNA single strand L1 is 29 bp; the length of DNA single strand L2 is 25 bp. In some other specific embodiments, the length of DNA single strand L1 is 29 bp; the length of DNA single strand L2 is 26 bp. In some other specific embodiments, the length of DNA single strand L1 is 29 bp; the length of DNA single strand L2 is 27 bp. In some other specific embodiments, the length of DNA single strand L1 is 29 bp; the length of DNA single strand L2 is 28 bp. In some other specific embodiments, the length of DNA single strand L1 is 29 bp; the length of DNA single strand L2 is 29 bp. In some other specific embodiments, the length of DNA single strand L1 is 29 bp; the length of DNA single strand L2 is 30 bp. In some other specific embodiments, the length of DNA single strand L1 is 29 bp; the length of DNA single strand L2 is 31 bp. In some other specific embodiments, the length of DNA single strand L1 is 29 bp; the length of DNA single strand L2 is 32 bp. In some other specific embodiments, the length of DNA single strand L1 is 29 bp; the length of DNA single strand L2 is 33 bp. In some other specific embodiments, the length of DNA single strand L1 is 29 bp; the length of DNA single strand L2 is 34 bp. In some other specific embodiments, the length of DNA single strand L1 is 29 bp; the length of DNA single strand L2 is 35 bp.

[0017] In some specific embodiments, the length of the single-stranded DNA L1 is 31 bp; the length of the single-stranded DNA L2 is 30 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 31 bp; the length of the single-stranded DNA L2 is 25 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 31 bp; the length of the single-stranded DNA L2 is 26 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 31 bp; the length of the single-stranded DNA L2 is 27 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 31 bp; the length of the single-stranded DNA L2 is 28 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 31 bp; the length of the single-stranded DNA L2 is 29 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 31 bp; the length of the single-stranded DNA L2 is 30 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 31 bp; the length of the single-stranded DNA L2 is 31 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 31 bp; the length of the single-stranded DNA L2 is 32 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 31 bp; the length of the single-stranded DNA L2 is 33 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 31 bp; the length of the single-stranded DNA L2 is 34 bp. In some other specific embodiments, the length of the single-stranded DNA L1 is 31 bp; the length of the single-stranded DNA L2 is 35 bp.

[0018] In some specific embodiments, the length of DNA single strand L1 is 32 bp; the length of DNA single strand L2 is 30 bp. In some other specific embodiments, the length of DNA single strand L1 is 32 bp; the length of DNA single strand L2 is 25 bp. In some other specific embodiments, the length of DNA single strand L1 is 32 bp; the length of DNA single strand L2 is 26 bp. In some other specific embodiments, the length of DNA single strand L1 is 32 bp; the length of DNA single strand L2 is 27 bp. In some other specific embodiments, the length of DNA single strand L1 is 32 bp; the length of DNA single strand L2 is 28 bp. In some other specific embodiments, the length of DNA single strand L1 is 32 bp; the length of DNA single strand L2 is 29 bp. In some other specific embodiments, the length of DNA single strand L1 is 32 bp; the length of DNA single strand L2 is 30 bp. In some other specific embodiments, the length of DNA single strand L1 is 32 bp; the length of DNA single strand L2 is 31 bp. In some other specific embodiments, the length of DNA single strand L1 is 32 bp; the length of DNA single strand L2 is 32 bp. In some other specific embodiments, the length of DNA single strand L1 is 32 bp; the length of DNA single strand L2 is 33 bp. In some other specific embodiments, the length of DNA single strand L1 is 32 bp; the length of DNA single strand L2 is 34 bp. In some other specific embodiments, the length of DNA single strand L1 is 32 bp; the length of DNA single strand L2 is 35 bp.

[0019] In some specific embodiments, the length of DNA single strand L1 is 33 bp; the length of DNA single strand L2 is 30 bp. In some other specific embodiments, the length of DNA single strand L1 is 33 bp; the length of DNA single strand L2 is 25 bp. In some other specific embodiments, the length of DNA single strand L1 is 33 bp; the length of DNA single strand L2 is 26 bp. In some other specific embodiments, the length of DNA single strand L1 is 33 bp; the length of DNA single strand L2 is 27 bp. In some other specific embodiments, the length of DNA single strand L1 is 33 bp; the length of DNA single strand L2 is 28 bp. In some other specific embodiments, the length of DNA single strand L1 is 33 bp; the length of DNA single strand L2 is 29 bp. In some other specific embodiments, the length of DNA single strand L1 is 33 bp; the length of DNA single strand L2 is 30 bp. In some other specific embodiments, the length of DNA single strand L1 is 33 bp; the length of DNA single strand L2 is 31 bp. In some other specific embodiments, the length of DNA single strand L1 is 33 bp; the length of DNA single strand L2 is 32 bp. In some other specific embodiments, the length of DNA single strand L1 is 33 bp; the length of DNA single strand L2 is 33 bp. In some other specific embodiments, the length of DNA single strand L1 is 33 bp; the length of DNA single strand L2 is 34 bp. In some other specific embodiments, the length of DNA single strand L1 is 33 bp; the length of DNA single strand L2 is 35 bp.

[0020] In some specific embodiments, the length of DNA single strand L1 is 34 bp; the length of DNA single strand L2 is 30 bp. In some other specific embodiments, the length of DNA single strand L1 is 34 bp; the length of DNA single strand L2 is 25 bp. In some other specific embodiments, the length of DNA single strand L1 is 34 bp; the length of DNA single strand L2 is 26 bp. In some other specific embodiments, the length of DNA single strand L1 is 34 bp; the length of DNA single strand L2 is 27 bp. In some other specific embodiments, the length of DNA single strand L1 is 34 bp; the length of DNA single strand L2 is 28 bp. In some other specific embodiments, the length of DNA single strand L1 is 34 bp; the length of DNA single strand L2 is 29 bp. In some other specific embodiments, the length of DNA single strand L1 is 34 bp; the length of DNA single strand L2 is 30 bp. In some other specific embodiments, the length of DNA single strand L1 is 34 bp; the length of DNA single strand L2 is 31 bp. In some other specific embodiments, the length of DNA single strand L1 is 34 bp; the length of DNA single strand L2 is 32 bp. In some other specific embodiments, the length of DNA single strand L1 is 34 bp; the length of DNA single strand L2 is 33 bp. In some other specific embodiments, the length of DNA single strand L1 is 34 bp; the length of DNA single strand L2 is 34 bp. In some other specific embodiments, the length of DNA single strand L1 is 34 bp; the length of DNA single strand L2 is 35 bp.

[0021] In some specific embodiments, the length of DNA single strand L1 is 35 bp; the length of DNA single strand L2 is 30 bp. In some other specific embodiments, the length of DNA single strand L1 is 35 bp; the length of DNA single strand L2 is 25 bp. In some other specific embodiments, the length of DNA single strand L1 is 35 bp; the length of DNA single strand L2 is 26 bp. In some other specific embodiments, the length of DNA single strand L1 is 35 bp; the length of DNA single strand L2 is 27 bp. In some other specific embodiments, the length of DNA single strand L1 is 35 bp; the length of DNA single strand L2 is 28 bp. In some other specific embodiments, the length of DNA single strand L1 is 35 bp; the length of DNA single strand L2 is 29 bp. In some other specific embodiments, the length of DNA single strand L1 is 35 bp; the length of DNA single strand L2 is 30 bp. In some other specific embodiments, the length of DNA single strand L1 is 35 bp; the length of DNA single strand L2 is 31 bp. In some other specific embodiments, the length of DNA single strand L1 is 35 bp; the length of DNA single strand L2 is 32 bp. In some other specific embodiments, the length of DNA single strand L1 is 35 bp; the length of DNA single strand L2 is 33 bp. In some other specific embodiments, the length of DNA single strand L1 is 35 bp; the length of DNA single strand L2 is 34 bp. In some other specific embodiments, the length of DNA single strand L1 is 35 bp; the length of DNA single strand L2 is 35 bp.

[0022] In the present invention, the complementary region of DNA single strand L1 and DNA single strand L2 is not less than 1 / 3 of the full length. For example, the complementary region of DNA single strand L1 and DNA single strand L2 is not less than 10 bases. In some embodiments, the complementary region is 10 bases, 11 bases, 12 bases, 13 bases, 14 bases, 15 bases, 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, 25 bases, 26 bases, 27 bases, 28 bases, 29 bases or 30 bases.

[0023] In the present invention, the non-complementary region of DNA single strand L1 and DNA single strand L2 is not less than 1 / 6 of the full length. For example, the complementary region of DNA single strand L1 and DNA single strand L2 is not less than 5 bases. In some embodiments, the complementary region is 5 bases, 6 bases, 7 bases, 8 bases, 9 bases, 10 bases, 11 bases, 12 bases, 13 bases, 14 bases or 15 bases.

[0024] In some specific embodiments:

[0025] The single-stranded L1 sequence is: 5’-Acrydite AAA AAA AAA AAA CCC TCT TCC TGG AAT TCC-3’;

[0026] The single-stranded L2 sequence is: 5’-Acrydite AAA ATA AAA AAA AAA AAA AAA AGGA ATT CC-3’;

[0027] In the present invention, the raw materials for preparing the hydrogel further include a cross-linking agent, and the cross-linking agent is at least one of N,N'-methylenebisacrylamide, polyethylene glycol 200, diacrylate (PEGDA-200), or polyethylene glycol 400 diacrylate (PEGDA-400).

[0028] In the present invention, the raw materials for preparing the hydrogel further include a photoinitiator, such as lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP), 1-hydroxycyclohexyl phenyl ketone (HCPK), etc.

[0029] In the present invention, the DNA hydrogel is driven by RNA as fuel. Its dynamic process includes the swelling reaction of RNA attacking the DNA double strand and the contraction behavior of the RNase H enzymatic reaction. Both start to act after RNA enters the hydrogel, and as time goes by, the reactions show different rates and the sol shows different volumes.

[0030] In some embodiments, the raw materials for preparing the hydrogel further include an RNA single strand R, which is complementary to the DNA single strand L1 or the DNA single strand L2.

[0031] Preferably, the length of the RNA single strand R is greater than the length of the complementary region between the DNA single strand L1 and the DNA single strand L2. However, the two L strands need to be longer than the RNA single strand R. Preferably, the length of the RNA single strand R is at least 1 base longer than the complementary region between the DNA single strand L1 and the DNA single strand L2. For example, the length of the RNA single strand R is 1 base, 2 bases, 3 bases, 4 bases, 5 bases, 6 bases, 7 bases, 8 bases, 9 bases, or 10 bases longer than the complementary region between the DNA single strand L1 and the DNA single strand L2.

[0032] In some embodiments, the length of the RNA single strand R is 6 bases longer than the complementary region between the DNA single strand L1 and the DNA single strand L2. In some specific embodiments, the nucleic acid sequence of the RNA single strand R is: 5’-CCA AUU CCA GGA AGA GGG-3’.

[0033] The present invention does not limit the sequences of single-stranded DNA or single-stranded RNA. The single-stranded DNA L1, single-stranded DNA L2, and / or single-stranded RNA R are only random sequences, and it is sufficient to meet the above conditions. They can also be sequences with therapeutic activity. For example, they can be RNAi sequences of disease treatment target genes, messenger ribonucleic acid (mRNA) sequences that can be transcribed into proteins, or ribosomal RNA (rRNA) sequences that can accelerate protein synthesis. The present invention does not limit this. The sequences with therapeutic activity can be the full lengths of single-stranded DNA L1, single-stranded DNA L2, and / or single-stranded RNA R, or only partial sequences. The present invention also does not limit this. As a feasible case, disease treatment target genes include, but are not limited to, EGFR (epidermal growth factor receptor), ALK (anaplastic lymphoma kinase), ROS1 (c-ros proto-oncogene), BRAF (B-type RAF kinase), MET (hepatocyte growth factor receptor, HGFR), HER2 (human epidermal growth factor receptor 2).

[0034] The hydrogel described in the present invention further includes ribonuclease and / or therapeutic agent.

[0035] In the present invention, the ribonuclease can degrade single-stranded RNA or RNA strands bound to DNA, but cannot degrade single-stranded DNA. For example, the ribonuclease is ribonuclease A, ribonuclease T1, and / or ribonuclease H.

[0036] In the present invention, the action form of the ribonuclease can be changed according to needs. Specifically, there are three types: a: without heating and without enzyme embedding, and there is only enzyme outside the gel; b: heating for enzyme embedding, but not performing secondary cleaning on the gel, and there is enzyme both inside and outside the gel; c: heating for enzyme embedding and cleaning the gel, and there is only enzyme inside the gel.

[0037] In the present invention, the therapeutic agent is a chemical drug, traditional Chinese medicine, or biological product, and the biological drug is a protein drug or nucleic acid drug. During the preparation of the gel, due to the lack of the presence of sodium and magnesium ions, the binding of DNA double strands is weak, thus opening the DNA double strands. Under such conditions, the gel is mixed with an aqueous solution of the drug, so that the drug can enter the interior of the gel. After standing for a period of time, sodium and magnesium ions are added to the solution (based on the concentration of sodium and magnesium ions in the RNase H reaction solution). After a period of time, the DNA strands restore the double helix structure to complete the loading of the therapeutic agent.

[0038] As a feasibility case, the chemical drugs include, but are not limited to, aspirin, aminophylline, adrenaline, morphine, metronidazole, cephalosporin, cimetidine, penicillin, roxithromycin, norfloxacin, acyclovir, ibuprofen, berberine, omeprazole, dexamethasone, chloramphenicol, ribavirin, metformin, atorvastatin, ciprofloxacin, oseltamivir, doxycycline, azithromycin, lidocaine.

[0039] The protein drugs include, but are not limited to, insulin, human growth hormone (hGH), erythropoietin (EPO), interferon (IFN), interleukin (IL), tumor necrosis factor (TNF), tissue plasminogen activator (tPA), coagulation factor VIII (FⅧ), coagulation factor IX (FⅨ), immunoglobulin (Ig), various antibodies, follicle-stimulating hormone (FSH), luteinizing hormone (LH), glucagon-like peptide-1 (GLP-1), octreotide and / or liraglutide;

[0040] The nucleic acid drugs include, but are not limited to, mRNA vaccines (such as the COVID-19 mRNA vaccine), ASO drugs (such as ASO drugs for treating spinal muscular atrophy (SMA), ASO drugs for treating hypercholesterolemia, etc.), small interfering RNA drugs (such as siRNA drugs for treating hereditary thyroxine-mediated amyloid polyneuropathy), nucleic acid aptamer drugs (such as aptamer drugs for treating wet age-related macular degeneration (AMD)).

[0041] Furthermore, the present invention also provides a preparation method of the hydrogel as described above, which includes mixing a water-soluble monomer, DNA single strand L1 and DNA single strand L2 with a crosslinking agent and then crosslinking to obtain the hydrogel.

[0042] During the crosslinking process, a crosslinking agent and / or a photoinitiator are also added. The conditions for crosslinking include, but are not limited to, ultraviolet irradiation. The crosslinking is carried out at room temperature, or can be carried out under heating conditions or can be carried out under conditions lower than room temperature, and the present invention does not make any limitation thereto.

[0043] The preparation method as described above further includes the step of adding ribonuclease and / or a therapeutic agent after swelling.

[0044] The precursor of the gel of the present invention presents a solution state before curing, while the solution becomes a gel state after curing, with an increase in viscosity and hardness. Further, the step of adding RNA single strand R is also included. The gel before curing is in a solution state, and adding RNA will not cause any change. After curing, adding RNA causes the gel to first expand and then return to its initial state.

[0045] The reaction process of the present invention first uses a permanent cross-linker Bis and a dynamic cross-linker DNA to polymerize through photoinitiation to finally form a three-dimensional network hydrogel. The dynamic process of the gel is achieved by RNA attacking the dynamic DNA cross-links and the RNase H enzymatic reaction. The dynamic process of the gel can be precisely controlled by the action form and dosage of RNase H and RNA to achieve dynamic control of the hydrogel strength. While the gel changes dynamically, the pore size inside the gel also changes, so the loading and release of materials can be carried out.

[0046] In some specific embodiments: The preparation method of the dynamic DNA hydrogel of the present invention includes:

[0047] 1) Self-assemble two complementary DNA single strands (L1 and L2) to form a DNA double helix structure.

[0048] 2) Dissolve a certain proportion of acrylamide, methylene bisacrylamide, DNA and LAP in an aqueous solution, and place the solution under ultraviolet light for photocuring.

[0049] 3) Swell the obtained gel in an aqueous solution.

[0050] 4) Place a certain amount of enzyme and the gel in a solution, heat to a certain temperature, keep warm for a certain time to allow the enzyme to enter the gel interior, and wash the gel after cooling.

[0051] 5) Place the gel in a solution and add a certain amount of RNA (R1) to form a dynamic hydrogel. The volume of the gel first increases and then returns to the initial state.

[0052] The DNA is a primer strand modified with methacrylamide and is artificially synthesized DNA.

[0053] Preferably, the concentration of each monomer is AAm: 1 - 4 mol / L, LAP: 0.5 mg - 3 mg / ml, Bis: 0.1 - 0.4% AAm, DNA: 0.1 - 0.3% AAm.

[0054] Preferably, in step 2, the gel is cured under the conditions of a photoinitiator and ultraviolet light irradiation.

[0055] Preferably, in step 2, a permanent cross-linker Bis and a drivable cross-linker (DNA cross-linker) are used to prepare a double-network three-dimensional network dynamic DNA hydrogel.

[0056] Preferably, the enzyme used in step 4 is selected as thermostable RNase H, the incubation temperature is 60 - 85 °C, and the incubation condition is 5 - 20 min.

[0057] Preferably, the DNA annealing process in step 1 is optimized as follows: Place equal amounts of DNA single strands L1 and L2 in the annealing solution, incubate at 95 °C for 5 min, and slowly cool to room temperature to ensure that L1 and L2 form a DNA double helix structure.

[0058] Preferably, the photoinitiator is LAP, and it can be replaced with photoinitiators of the same type such as 1-hydroxycyclohexyl phenyl ketone (HCPK), etc.

[0059] In step 4, the enzyme and the gel are heated to achieve enzyme entrapment, and the enzyme dosage is 0.5 - 4 U / ul. Preferably, the enzyme entrapment conditions are as follows: Use RNase H reaction buffer as the solution for enzyme entrapment.

[0060] Preferably, in step 3, the prepared DNA gel is placed in an aqueous solution for maximum swelling, and residual monomers and initiators are removed. Among them, the maximum swelling time of the gel is 12 - 24 h.

[0061] Preferably, in step 5, the ratio of the amount of RNA added to the DNA in the gel is 1:1.

[0062] In step 5, the DNA gel is driven by RNA as fuel. Its dynamic process includes the swelling reaction of RNA attacking the DNA double strand and the contraction behavior of the RNase H enzymatic reaction. Both start to act after RNA enters the gel, and as time goes by, the reactions exhibit different rates, and the sol shows different volumes.

[0063] Furthermore, the present invention also provides the application of the hydrogel as described above and / or the hydrogel prepared by the preparation method as described above in the preparation of drugs and / or drug carriers.

[0064] Even further, the present invention also provides drugs and / or drug carriers, including the hydrogel as described above and / or the hydrogel prepared by the preparation method as described above.

[0065] The gel provided by the present invention can achieve the change from liquid to gel peptide, and can also achieve the attachment or release of drugs with the change of internal pore size. Therefore, it can be used as a drug itself or as a drug carrier. As a feasible case, it can be used as a medical adjuvant to promote the healing of wounds in the body. It can also be used to load insulin to achieve a sustained release effect. In addition, the gel as described above can also be used as a cell culture medium, for example, for culturing human cells, animal cells or microorganisms.

[0066] Even further, the present invention also provides a treatment method, which includes administering the drugs and / or drug carriers as described above.

[0067] In the described treatment method, only the drug as described above may be administered, or only the drug encapsulated by the drug carrier as described above may be administered, or both may be administered.

[0068] In the case of administering both, two or more drugs may be administered simultaneously or sequentially, and the present invention does not limit this. For example, the administration interval between the two drugs is not longer than 6 hours, preferably not longer than 5 hours, more preferably not longer than 4 hours, even more preferably not longer than 3 hours, still more preferably not longer than 2 hours, and most preferably not longer than 1 hour. The administration frequencies of the two drugs in the drug may be the same or different, and the present invention does not limit this. For example, the administration frequencies of the two drugs in the drug are independently selected from 4 times / day, 3 times / day, 2 times / day, 1 time / day, once every 2 days, once every 3 days, once every 5 days, once every 7 days, once every 10 days, once every 20 days, or once every 30 days.

[0069] In the treatment method as described above, the subject to be administered is a human or an animal, and the animal is a mammal. As a feasible case, it may be a feline, a canine, a rodent, a primate, a bird, a bovine, a horse, and / or a porcine.

[0070] The present invention uses a permanent cross-linking agent and a dynamic cross-linking agent DNA to polymerize by a photo-initiated method to finally form a three-dimensional network-structured hydrogel. The dynamic process of the gel is realized by RNA attacking the dynamic DNA cross-linking points and the RNase H enzymatic reaction. The dynamic process of the gel can be precisely controlled by the action form and dosage of RNase H and RNA, so as to realize the dynamic control of the strength of the hydrogel. While the gel is dynamically changing, the pore size inside the gel is also changing, so the loading and release of materials can be carried out. DNA, RNA, and RNase H have good biocompatibility and can be used as medical dressings; the process of preparing the hydrogel is simple and convenient, and it can be applied to fields such as medical dressings, drug sustained release, and cell-free protein synthesis. Description of the Drawings

[0071] Figure 1 The DNA hydrogel prepared in Example 1 and its swelling in solution;

[0072] Figure 2 Photos of the RNA-driven DNA hydrogel at different times;

[0073] Figure 3 The cyclic thermal-driven size change curve of the DNA hydrogel;

[0074] Figure 4 The dynamic change diagram of the diameter of the DNA hydrogel prepared in Example 2;

[0075] Figure 5 The dynamic change diagram of the diameter of the DNA hydrogel prepared in Comparative Example 1;

[0076] Figure 6 . Relationship curve between adriamycin concentration and drug loading amount in Example 3;

[0077] Figure 7 . Relationship curve between HeLa cell survival rate and RNA / DNA ratio in Example 4. Specific implementation manners

[0078] The present invention provides a dynamic DNA hydrogel and a preparation method thereof. Those skilled in the art can draw on the content of this article and appropriately modify process parameters to achieve it. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all regarded as included in the present invention. The methods and applications of the present invention have been described through preferred embodiments. It is obvious that relevant personnel can make changes or appropriate changes and combinations to the methods and applications in this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0079] Unless otherwise defined in the present invention, scientific and technical terms related to the present invention shall have the meanings understood by those of ordinary skill in the art.

[0080] In addition, unless otherwise specified in this article, terms in the singular form in this article shall include the plural form, and terms in the plural form shall include the singular form. More specifically, as used in this specification and the appended claims, unless otherwise clearly indicated, the singular forms "a" and "the" include plural referents.

[0081] The terms "comprising", "including" and "having" can be used interchangeably, aiming to indicate the inclusiveness of the solution, meaning that the solution may have other elements in addition to the listed elements. At the same time, it should be understood that when using "comprising", "including" and "having" to describe in this article, the solution of "consisting of..." is also provided.

[0082] When the term "and / or" is used in this article, it includes the meanings of "and", "or" and "any other combination of all or part of the elements linked by the term".

[0083] The numerical ranges and parameters involved in the present invention have been presented as precisely as possible in the specific embodiments. However, any value inevitably contains standard deviations caused by individual test methods. Therefore, unless otherwise clearly stated, it should be understood that all numerical ranges or specific data used in this disclosure may have certain reasonable deviations within a certain range, for example: within ±10%, ±5%, ±1% or ±0.5%.

[0084] The test materials used in the present invention are all ordinary commercially available products and can be purchased in the market. The present invention will be further described below in conjunction with embodiments:

[0085] Example 1.

[0086] Self-assemble single-stranded DNA to form a DNA double helix structure. First, configure the specific parameters of the DNA annealing solution as follows: 10 mM Tris-HCl (pH 8.0), 50 mM NaCl). Dissolve 100 nmol of L1 and L2 in 5 μl of the DNA annealing solution respectively, and then mix the two and shake well. Seal the mixed solution and heat it at 95 °C for 5 min, and slowly cool it to room temperature (>2 h) to obtain a DNA solution with a concentration of 10 nmol / μl. The sequence of L1 is: 5’-Acrydite AAA AAA AAA AAA CCC TCT TCC TGGAAT TCC-3’. The sequence of L2 is: 5’-Acrydite AAA ATA AAA AAA AAA AAA AAA AGGA ATT CC-3’.

[0087] 2) Dissolve a certain amount of AAm in water to obtain an AAm solution with a concentration of 9 mol / L of AAm, and add Bis (0.1% of AAm, molar ratio), DNA (0.1% of AAm, molar ratio), and LAP (0.5 mg / ml) to the AAm solution. Vortex and shake well.

[0088] 3) Drop the prepared mixed solution (1-2 μl) onto a super-hydrophobic treated glass substrate to form a spherical droplet, and place it under ultraviolet light for reaction for 20 min to obtain a gel. Immerse the gel in the solution for 24 h to swell it to the maximum.

[0089] 4) Place the swollen gel in the solution, add RNase H enzyme, and calculate the amount of the enzyme according to 2 U / μl to prepare a solution. Heat it to 80 °C, keep it warm for 15 min, then cool it at room temperature for 2 min, and then cool it at 2-8 °C for 20 min. Complete the embedding of RNase H enzyme. And wash it with the RNase H enzyme reaction solution. Complete the primary preparation of the dynamic DNA gel.

[0090] 5) Place the gel in 20 μl of the RNase H enzyme reaction solution, and add RNA with a dosage of 1 times the amount of DNA to complete the preparation of the dynamic DNA gel. The gel first swells and then returns to its initial state. The RNA sequence R1 is: 5’-GGA AUU CCA GGA AGA GGG-3’.

[0091] Example 2.

[0092] 1) Self-assemble single-stranded DNA to form a DNA double helix structure. First, configure the specific parameters of the DNA annealing solution as follows: 10 mM Tris-HCl (pH 8.0), 50 mM NaCl. Dissolve 100 nmol of L3 and L4 in 5 μl of the DNA annealing solution respectively, and then mix the two and shake well. Seal the mixed solution and heat it at 95 °C for 5 min, and then slowly cool it to room temperature (>2 h) to obtain a DNA solution with a concentration of 10 nmol / μl. The sequence of L3 is: 5’-Acrydite AAA AA CCC TTC CGG GCC CTTCCC-3’. The sequence of L4 is: 5’-Acrydite AAA AA GGG AAG CGG C-3’.

[0093] 2) Dissolve a certain amount of AAm in water, and add Bis (0.1% AAM), DNA (0.1% AAM) and LAP (0.5 mg / ml) to the AAm solution. Vortex and shake well.

[0094] 3) Drop the prepared mixed solution (1 - 2 μl) onto the super-hydrophobic treated glass substrate to form spherical droplets, and place them under ultraviolet light for 20 min to obtain a gel. Immerse the gel in the solution for 24 h to swell it to the maximum.

[0095] 4) Place the swollen gel in the solution, add RNase H enzyme, and calculate the dosage of the enzyme at 2 U / μl to prepare a solution. Heat it to 80 °C, keep it warm for 15 min, then cool it at room temperature for 2 min, and then cool it at 2 - 8 °C for 20 min. Complete the embedding of RNase H enzyme. And wash it with the RNase H enzyme reaction solution. Complete the primary preparation of the dynamic DNA gel.

[0096] 5) Place the gel in 20 μl of the RNase H enzyme reaction solution, and add RNA with a dosage 1 times that of DNA to complete the preparation of the dynamic DNA gel. The gel first swells and then returns to its initial state. The sequence of RNA is R2: 5’-GGG AAG GGC CCG GAAGGG-3’.

[0097] Comparative Example 1

[0098] 1) Self-assemble single-stranded DNA to form a DNA double helix structure. First, configure the specific parameters of the DNA annealing solution: 10 mM Tris-HCl (pH 8.0), 50 mM NaCl. Dissolve 100 nmol of L3 and L4 in 5 μl of the DNA annealing solution respectively, and then mix and shake well. Seal the mixed solution and heat it at 95 °C for 5 min, and slowly cool it to room temperature (>2 h) to obtain a DNA solution with a concentration of 10 nmol / μl. The sequence of L3 is: 5’-Acrydite AAA AA CCC TTC CGG GCC CTTCCC-3’. The sequence of L4 is: 5’-Acrydite AAA AA GGG AAG GGC C-3’.

[0099] 2) Dissolve a certain amount of AAm in water, and add Bis (0.1% AAM), DNA (0.1% AAM) and LAP (0.5 mg / ml) to the AAm solution. Vortex and shake well.

[0100] 3) Drop the prepared mixed solution (1 - 2 μl) onto the superhydrophobic-treated glass substrate to form a spherical droplet, and place it under ultraviolet light for reaction for 20 min to obtain a gel. Immerse the gel in the solution for 24 h to swell it to the maximum.

[0101] 4) Place the swollen gel in the solution, and add RNase H enzyme. The dosage of the enzyme is calculated as 1 U / μl to prepare a solution. Heat it to 80 °C, keep it warm for 15 min, then cool it to room temperature for 2 min, and cool it at 2 - 8 °C for 20 min. Complete the embedding of RNase H enzyme. And wash it with the RNase H enzyme reaction solution. Complete the primary preparation of the dynamic DNA gel.

[0102] 5) Place the gel in 20 μl of the RNase H enzyme reaction solution, and add RNA with a dosage 1 times that of DNA to complete the preparation of the dynamic DNA gel. The gel first swells and then returns to its initial state. The RNA sequence is R2: 5’-GGG AAG GGC CCG GAAGGG-3’.

[0103] Effect detection:

[0104] The gel as described above can be characterized by infrared spectroscopy. Its main change is that there are a large number of double bonds in the solution before curing. There is an infrared characteristic absorption peak of double bonds (1640 cm-1) in the infrared spectrum. And the characteristic absorption peak disappears after curing.

[0105] Viscosity and hardness judgment: Before curing, the precursor presents a solution state, while after curing, the solution becomes a gel state, with an increase in viscosity and hardness. A rheometer, TMA, and DMA can be used to test the viscosity modulus.

[0106] Gel-driven changes: Before curing, the gel is in a solution state, and adding RNA will not cause any changes. After curing, when RNA is added, the gel first swells and then returns to its initial state.

[0107] Some of the results are as follows:

[0108] After testing, the gel properties of Examples 1-2 are different. It is analyzed that this is due to the different formulations used. For example, the different lengths of DNA, the different amounts of Bis, DNA, and enzymes. The gels of Examples 1-2 each have their own advantages. It was found in the experiment that the longer the DNA length, the longer the uncomplemented part, the softer the gel, and the greater the gel swelling when RNA is added for driving. The greater the amount of Bis used, the harder the gel, and the corresponding lower swelling rate when RNA is added for driving. The more enzyme used, the faster the RNA hydrolysis, and the faster the gel driving cycle is completed. Correspondingly, the smaller the swelling.

[0109] In addition, as Figure 5 shown, the experiment was carried out on the basis of Example 2, and the difference lies in Figure 5 the amount of enzyme used to prepare the gel in [[]] is 1 U / μl. In Comparative Example 1, the gel can swell during driving, but it fails to return to its initial state, indicating that under some parameters, the performance of the gel cannot meet the use standard.

[0110] Comparative Example 2

[0111] The other step parameters are the same as those in Example 1, but the amount of BIS used is too much (when it exceeds 0.4% of the AAm amount), and the RNase H enzyme cannot enter the interior of the gel, and the gel can only swell but cannot shrink.

[0112] Comparative Example 3

[0113] The other step parameters are the same as those in Example 1, but the amount of BIS used in step 2) is too little (when the amount of bis is less than 0.1% of AAm), and there are too few gel fixed crosslinking points. When the DNA crosslinking points are opened, the gel is too soft, which may lead to deformation and collapse, etc., and the gel cannot return to its initial state when it shrinks.

[0114] Comparative Example 4

[0115] The other step parameters are the same as those in Example 1, but the enzyme embedding temperature in step 4) exceeds 90 °C and the time is long, which will cause the enzyme to be inactivated, so that the gel cannot shrink.

[0116] Example 3

[0117] Refer to the steps of Example 1 to prepare the DNA dynamic hydrogel. Place the DNA dynamic hydrogel in a certain amount of distilled water for swelling, open the DNA double strands to make the pore size of the DNA gel larger, add different amounts of doxorubicin solution for drug embedding, let it stand in the dark for 48 h for drug loading, then add a certain amount of sodium chloride and magnesium chloride to the solution, let it stand for 12 hours to close the DNA double strands, collect the supernatant, and calculate the drug loading amount using the concentration difference.

[0118] Example 4

[0119] Refer to the steps of Example 3 to prepare the drug-loaded DNA dynamic hydrogel. Dilute the HeLa cell suspension according to the designed concentration and inoculate it into a 96-well culture plate, and place it in an incubator for adherent culture. After the cells adhere, perform the drug addition treatment with the DNA dynamic hydrogel, and realize drug release by the drive of RNA. The RNA:DNA ratio gradients are 0.4, 0.6, 0.8, 1, and 1.2. Culture the cells with the drug for 24 h, then add MTT and continue to culture for 4 h. Aspirate the culture medium, add DMSO, shake the plate using a microplate reader, and detect the absorbance at 492 nm to calculate the cell survival rate.

[0120] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. Dynamic DNA hydrogel, the raw materials for its preparation include: Water-soluble monomer, DNA single strand L1 and DNA single strand L2; The DNA single strand L1 is partially complementary to the DNA single strand L2; The ends of the DNA L1 and the DNA single strand L2 are modified with polymerizable functional groups respectively.

2. The hydrogel according to claim 1, characterized in that The complementary region between DNA single strand L1 and DNA single strand L2 is not less than 10 bases; the non-complementary region is not less than 5 bases; The polymerizable functional group is an acrylic water-soluble monomer, preferably at least one of acrylamide, acrylic acid, 2-acrylamido-2-methyl-1-propane sulfonic acid, hydroxyethyl acrylate, acryloyl glycinamide or methacrylic acid.

3. The hydrogel according to claim 1 or 2, characterized in that The preparation raw materials also include a cross-linking agent and / or an initiator; The initiator is lithium phenyl-2,4,6-trimethylbenzoylphosphinate or 1-hydroxycyclohexyl phenyl ketone; The cross-linking agent is at least one of methylene bisacrylamide, polyethylene glycol 200, diacrylate or polyethylene glycol 400 diacrylate.

4. The hydrogel according to any one of claims 1 to 3, characterized in that It also includes an RNA single strand R, which is complementary to the DNA single strand L1 or the DNA single strand L2; preferably, the length of the RNA single strand R is greater than the length of the complementary region between the DNA single strand L1 and the DNA single strand L2; preferably, the length of the RNA single strand R is at least 4 bases longer than the complementary region between the DNA single strand L1 and the DNA single strand L2.

5. The hydrogel according to any one of claims 1 to 4, characterized in that The DNA single strand L1, the DNA single strand L2 and / or the RNA single strand R have therapeutic activity.

6. The hydrogel according to any one of claims 1 to 5, characterized in that Also included are ribonucleases and / or therapeutic agents.

7. The method for preparing the hydrogel according to any one of claims 1 to 6, comprising mixing a water-soluble monomer, a DNA single strand L1 and a DNA single strand L2 with a crosslinking agent, and crosslinking to obtain a hydrogel.

8. The preparation method according to claim 7, characterized in that: Also included is the step of adding ribonuclease and / or therapeutic agent after swelling; Furthermore, the method further comprises the step of adding RNA single-strand R.

9. Use of the hydrogel according to any one of claims 1 to 6 and / or the hydrogel prepared by the preparation method according to claim 7 or 8 in the preparation of drugs and / or drug carriers.

10. A drug and / or a drug carrier, comprising the hydrogel according to any one of claims 1 to 6 and / or the hydrogel prepared by the preparation method according to claim 7 or 8.

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