Search

CN-118240813-B - Biomimetic mineralization material with high stability for protecting DNA and preparation method thereof

CN118240813BCN 118240813 BCN118240813 BCN 118240813BCN-118240813-B

Abstract

The invention discloses a biomimetic mineralization material with high stability for protecting DNA and a preparation method thereof. The method comprises the steps of self-assembling DNA molecules and divalent metal ions to obtain DNA/metal nano-particles, wrapping polyelectrolyte layers on the surfaces of the DNA/metal nano-particles through a layer-by-layer assembly technology to obtain DNA/metal@LBL nano-particles, depositing polydopamine layers on the surfaces of the DNA/metal@LBL nano-particles to obtain DNA/Fe@LBL@PDA particles, and thus the biomimetic mineralizing material with high stability for protecting DNA is obtained. The biomimetic mineralized material has the advantages of simple construction, high density, long-acting stability and the like, and can effectively protect the stability and the integrity of DNA molecules in the environment of severe external storage (such as ROS, high temperature, high salt, alkalinity, humidity, biological solvent, nuclease and the like). The invention can provide technical support for DNA high-density storage and long-term stability storage, and has wide application value in the fields of biological medicine, genetic engineering, disease treatment and DNA storage.

Inventors

  • LU CHUNHUA
  • ZHONG WUKUN
  • YANG HUANGHAO

Assignees

  • 福州大学

Dates

Publication Date
20260508
Application Date
20240308

Claims (6)

  1. 1. A preparation method of a biomimetic mineralization material with high stability protection DNA is characterized by comprising the following steps: 1) Self-assembling the DNA molecules with divalent metal ions to obtain DNA/metal nano-particles; 2) Coating a polyelectrolyte layer on the surface of the DNA/metal nano-particles by a layer-by-layer assembly technology to obtain the DNA/metal@LBL nano-particles, namely adding polycation electrolyte into the DNA/metal nano-particle solution for reaction, adding polyanion electrolyte, and continuing the reaction so as to coat the polyelectrolyte layer on the surface of the DNA/metal nano-particles to obtain the DNA/metal@LBL nano-particle solution; 3) Depositing a polydopamine layer on the surface of the DNA/metal@LBL nano-particles to obtain DNA/Fe@LBL@PDA particles, namely the biomimetic mineralization material with high stability for protecting the DNA; the divalent metal ion is ferrous ion; the polycation electrolyte is polyallylamine hydrochloride; the polyanionic electrolyte is poly (4-styrenesulfonic acid).
  2. 2. The preparation method of claim 1, which is characterized by comprising the following specific steps: 1) Preparing a DNA molecule into a DNA molecule solution with the concentration of 1-10 mu M by using enzyme-free water, preparing a salt compound of divalent metal ions into a divalent metal ion salt solution with the concentration of 1-20 mM by using enzyme-free water, uniformly mixing the DNA molecule solution and the divalent metal ion salt solution in deionized water, placing the mixture in a 95 ℃ metal bath for incubation for 0.5-3 h, and naturally cooling to room temperature to obtain a DNA/metal nanoparticle solution; 2) Adding polycation electrolyte into the DNA/metal nanoparticle solution, reacting for 10-60 min at room temperature, adding polyanion electrolyte, and continuing reacting for 10-60 min at room temperature so as to wrap a polyelectrolyte layer on the surface of the DNA/metal nanoparticle to obtain the DNA/metal@LBL nanoparticle solution; 3) Adding dopamine monomer into the DNA/metal@LBL nanoparticle solution, adjusting the pH to 8.5, placing the mixture in an oscillator, and reacting for 2-16 hours at room temperature and the rotating speed of 100-700 rpm so as to deposit a polydopamine layer on the surface of the DNA/metal@LBL nanoparticle to obtain the DNA/Fe@LBL@PDA particle solution; 4) And (3) carrying out centrifugal washing and purification treatment on the DNA/Fe@LBL@PDA particle solution by adopting ultrapure water, separating to obtain purified DNA/Fe@LBL@PDA particles, namely the biomimetic mineralized material with high stability for protecting the DNA, and carrying out vacuum freeze drying and preservation.
  3. 3. The method of claim 2, wherein the DNA molecule comprises, but is not limited to, one or more double-stranded DNA molecules, single-stranded DNA molecules, and circular DNA molecules, with or without modification.
  4. 4. The method of claim 2, wherein the centrifugal washing is performed at 8000-12000 rpm for 5-20 min and 3-6 times.
  5. 5. A biomimetic mineralizing material with high stability and protection of DNA, which is characterized by being obtained by the preparation method of claim 1.
  6. 6. The preparation method of claim 1 or the application of the biomimetic mineralizing material of claim 5 in the field of DNA storage.

Description

Biomimetic mineralization material with high stability for protecting DNA and preparation method thereof Technical Field The invention belongs to the field of synthetic biological material science, and particularly relates to a biomimetic mineralization material with high stability for protecting DNA and a preparation method thereof. Background DNA is genetic material in organisms, has extremely high storage density, and is a natural carrier for bearing various life genetic information and disease information. DNA is widely used in the fields of drug delivery, genetic engineering, DNA-based structural design, and ideal information storage media due to its ease of modification, precise assembly, efficient disease treatment, and high storage density. However, DNA molecules are extremely vulnerable to damage by nucleases, metal ions, oxidative free radicals, and high temperatures in the environment, under unprotected conditions, leading to the loss of information and functions carried by themselves. Methods for protecting DNA molecules are currently known, mainly desiccation, cryopreservation, chemical modification, silica mineralization, etc. However, these methods are still not capable of long-term protection of DNA molecules, and have the disadvantages of high maintenance costs or expensive and toxic reagents used in the preparation process. In view of the above, development of a biomimetic mineralized material which is environment-friendly, low in cost and capable of protecting DNA with high stability is expected to realize long-term stability and integrity of the DNA in the long-term storage process. Disclosure of Invention The invention aims to provide a biomimetic mineralization material with high stability for protecting DNA and a preparation method thereof, which can improve the stability, integrity and effectiveness of DNA molecules under complex storage conditions. The technical scheme adopted by the invention is as follows: A preparation method of a biomimetic mineralization material with high stability protection DNA comprises the following steps: 1) Self-assembling the DNA molecules with divalent metal ions to obtain DNA/metal nano-particles; 2) Coating a polyelectrolyte layer on the surface of the DNA/metal nano-particles by a layer-by-layer assembly technology to obtain the DNA/metal@LBL nano-particles; 3) Depositing a polydopamine layer on the surface of the DNA/metal@LBL nano particle to obtain the DNA/Fe@LBL@PDA particle, namely the biomimetic mineralization material with high stability for protecting the DNA. Furthermore, the preparation method of the biomimetic mineralization material with high stability protection DNA comprises the following specific steps: 1) Preparing a DNA molecule into a DNA molecule solution with the concentration of 1-10 mu M by using enzyme-free water, preparing a salt compound of divalent metal ions into a divalent metal ion salt solution with the concentration of 1-20 mM by using enzyme-free water, uniformly mixing the DNA molecule solution and the divalent metal ion salt solution in deionized water, placing the mixture in a 95 ℃ metal bath for incubation for 0.5-3 h, and naturally cooling to room temperature to obtain a DNA/metal nanoparticle solution; 2) Adding polycation electrolyte into the DNA/metal nanoparticle solution, reacting for 10-60 min at room temperature, adding polyanion electrolyte, and continuing reacting for 10-60 min at room temperature so as to wrap a polyelectrolyte layer on the surface of the DNA/metal nanoparticle to obtain the DNA/metal@LBL nanoparticle solution; 3) Adding dopamine monomer into the DNA/metal@LBL nanoparticle solution, adjusting the pH to 8.5, placing the mixture in an oscillator, and reacting for 2-16 hours at room temperature and the rotating speed of 100-700 rpm so as to deposit a polydopamine layer on the surface of the DNA/metal@LBL nanoparticle to obtain the DNA/Fe@LBL@PDA particle solution; 4) Carrying out centrifugal washing and purification treatment on the DNA/Fe@LBL@PDA particle solution by adopting ultrapure water, separating to obtain purified DNA/Fe@LBL@PDA particles, namely the biomimetic mineralized material with high stability protection DNA, and carrying out vacuum freeze drying preservation; wherein the DNA molecules include, but are not limited to, one or more double-stranded DNA molecules, single-stranded DNA molecules, circular DNA molecules, with or without modification; wherein the divalent metal ions include, but are not limited to, ferrous ions, cupric ions, and zinc ions; wherein the polycationic electrolyte includes, but is not limited to, polyallylamine hydrochloride, polyethylenimine, polydimethyldiallylammonium chloride; The polyanion electrolyte comprises, but is not limited to, poly (4-styrenesulfonic acid), polyacrylic acid and sodium polystyrene sulfonate, wherein the rotational speed of centrifugal washing is 8000-12000 rpm, the duration is 5-20 min, and the times are 3-6. A biomimetic m