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Nucleic acid self-assembled composite nanoflower particle material as well as preparation method and application thereof

A composite material and self-assembly technology, applied in chemical instruments and methods, pharmaceutical formulations, other chemical processes, etc., can solve problems such as reducing meaning

Active Publication Date: 2017-06-13
SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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  • Abstract
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Problems solved by technology

[0005] However, it is a good idea to prepare nano-medicines by using constant temperature rolling circle amplification reaction to prepare DNA macromolecules as a carrier. The research on this aspect has just started, and there are still many problems to be solved and improved; for example: (1) people have obtained DNA nanoparticles are the direct product of the constant temperature rolling circle amplification reaction. Although the mechanism of its formation has been discussed to some extent, how to controllably prepare such nanoparticles and effectively regulate their size and shape is still a problem. Difficulties; (2) This type of DNA nanoparticles still cannot directly pass through the cell membrane, and needs to be modified with a layer of positively charged polymers on its surface. This design greatly reduces the significance of using DNA macromolecules as drug carriers; (3) Select and DNA-like biomaterials to modify and stabilize DNA nanoparticles is another problem that needs to be solved

Method used

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  • Nucleic acid self-assembled composite nanoflower particle material as well as preparation method and application thereof
  • Nucleic acid self-assembled composite nanoflower particle material as well as preparation method and application thereof
  • Nucleic acid self-assembled composite nanoflower particle material as well as preparation method and application thereof

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Effect test

Embodiment 1

[0056] Embodiment 1 prepares self-assembled composite material

[0057] The preparation method of the self-assembled composite material is as follows figure 1 shown, including the following steps:

[0058] (1) Amplify the nucleic acid, specifically using the rolling circle amplification technique. Specifically, the linear single-stranded template DNA and the primer (an oligonucleotide complementary to the template sequence) recognize each other, and the two ends of the first and last ends are closed to form a circle. Linked into circular single-stranded DNA under the action of ligase;

[0059] (2) In the presence of polymerase and dNTPs, copy the circular template from the primer, perform infinite single-stranded amplification through strand displacement, and generate long single-stranded nucleic acid molecules with repeating template sequences;

[0060] (3) Prepare a DNA aqueous solution with a mass concentration of 0.1 mg / ml, and add Mg to the DNA aqueous solution 2+ solu...

Embodiment 2

[0064] In this example, the self-assembled composite material of Example 1 and magnesium pyrophosphate were stained with gel red, and verified with a confocal laser scanning microscope, and the results are shown in FIG. 2 .

[0065] from Figure 2(a)-2(b) It can be seen that when excited under 405nm excitation light, Mg 2 PPi / DNA-NFs emit distinct blue fluorescence, while Mg 2 P 2 o 7 The solution still cannot observe obvious blue fluorescence under a larger magnification. Gel red can stain DNA molecules, but it cannot stain inorganic crystals. Observation by confocal laser scanning microscope (CLSM) confirms that DNA molecules are wrapped in in nanoflower particles.

Embodiment 3

[0067] This embodiment verifies the self-assembled composite material of embodiment 1 with TGA and XRD, and the results are as follows image 3 shown.

[0068] Depend on image 3 It can be seen from the TGA diagram that: Mg 2 The weight loss of PPi / DNA-NFs is divided into three stages: 50-137℃, 6.5% weight loss, which is the volatilization of free water; DNA begins to pyrolyze; 183-800°C, 22.7% weight loss, this part is the pyrolysis of DNA, 58.5% mass remains in the whole thermal process, due to Mg 2 The melting point of PPi is 1383°C, so the residual substance at this stage is Mg 2 PPi, indicating that the prepared composite material has both DNA and Mg 2 Components of PPi.

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Abstract

The invention belongs to the technical field of biology and relates to a nucleic acid self-assembled composite nanoflower particle material as well as a preparation method and application thereof. The self-assembled composite material refers to nanoflower particles formed by cocrystallizing contracted nucleic acid and Mg2P2O7, wherein the particle size of the particles is less than 500nm. The self-assembled composite material disclosed by the invention is of a nanoflower structure with high specific surface area, the DNA nanoflower self-assembled composite material has the yield close to 100% and the particle size of 500nm or less, and later utilization is facilitated; moreover, the DNA or RNA in the self-assembled composite material disclosed by the invention is an endogenous macromolecule in animal cells and has low toxicity and excellent biocompatibility, the DNA serves as a carrier of genetic information and can also achieve effects of targeting identification, enzyme catalysis and the like, multifunctional integration of the DNA nanoflower composite material can be realized by introducing functional nucleic acids, natural bio-molecules are totally adopted, and the self-assembled composite material is high in biocompatibility, low in cytotoxicity, low in production cost and favorable or large-scale production and development.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a self-assembled composite material and its preparation method and application, in particular to a nucleic acid self-assembled composite nano flower particle material and its preparation method and application. Background technique [0002] Cancer is currently the number one disease threatening human life and health. Fundamentally speaking, cancer is a complex genetic disease, and DNA is the most important biological macromolecule, which is naturally selected to store biological genetic information. Due to the highly specific Watson-Crick base interaction, in the 1990s, Professor Seeman of New York University discovered that two-dimensional and three-dimensional nanostructures could be constructed by using DNA controllable self-assembly, which opened the development of DNA nanotechnology. [0003] As a nano drug carrier, DNA macromolecules have advantages unmatched by other technologi...

Claims

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Application Information

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IPC IPC(8): B01J20/24B01J20/30A61K47/26A61K9/14
CPCA61K9/145B01J20/048B01J20/24B01J2220/4856
Inventor 田雷蕾袁雪霞邵晨赵浩然徐婧媛
Owner SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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