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A kind of two-component gene carrier and its application

A gene carrier, two-component technology, applied in the field of gene carrier

Inactive Publication Date: 2011-12-14
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As far as we know, there are no literature or patent reports on the synthesis of brush-type anionic block copolymers based on polyethylene glycol methacrylate containing mercapto groups (-SH), the cross-linking of disulfide bonds, and their Application to the construction of composite gene carriers with magnetic responsiveness and protein adsorption resistance

Method used

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  • A kind of two-component gene carrier and its application
  • A kind of two-component gene carrier and its application
  • A kind of two-component gene carrier and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Example 1: Synthetic method of polycationic magnetic nanoparticles

[0078] (1) Synthetic method of cationic polymer

[0079] Put the vial and syringe equipped with a stirrer in an oven at 120 ℃ to dry overnight, take out the vial and plug it with a ground glass stopper, and cool it with a pump until it reaches room temperature. Pure argon. Inject a certain amount of anhydrous tetrahydrofuran (THF) that has been refluxed to remove water and dried into the branch bottle containing KH powder, stir and wash, and suck out the THF containing mineral oil with a syringe after resting, repeat this three times, and finally use high-purity The residual THF solution was blown dry with argon. Inject a certain amount of THF; place the reaction bottle in an oil bath at 45 °C, stir it magnetically, and at the same time inject cholesterol equal to half the amount of KH substance (pre-dissolved in anhydrous THF, protected by argon, and cholesterol is abbreviated as Chol- ), initiate ...

Embodiment 2

[0083] Example 2: Brush type diblock copolymer PPEGMA 24 - b -P t BMA 49 synthesis method

[0084] (1) ATRP macroinitiator PPEGMA 24 Preparation and purification of -Cl: Put the vial in an oven at 120°C to dry in advance, take it out, put a ground glass stopper on it, and cool it with a pump until it reaches room temperature. Number average molecular weight 300 g mol -1 The monomer polyethylene glycol methacrylate (PEGMA) (2.6 g, 8.80 mmol) was dissolved in ethanol / water (95 / 5 v / v; 8.75 mL), added to the vial, and the initiator α-bromo Methyl propionate (50 mu L, 0.440 mmol), ligand 2,2′-bipyridine (138.9 mg, 0.880 mmol), cuprous chloride (44.0 mg, 0.440 mmol) with a purity of 99.99% were added to the above mixed system, and the high-purity Argon protection. Raise the temperature to 30°C, and maintain this temperature, stir with magnetic force, and react for about 10-12 hours.

[0085] The reaction product was diluted by adding absolute ethanol, and contacted with ...

Embodiment 3

[0090] Example three: brush block copolymer PPEGMA 32 - b -P t BMA 53 synthesis method

[0091] (1) ATRP macroinitiator PPEGMA 32 Preparation and purification of -Cl: Put the vial in an oven at 120°C to dry in advance, take it out, put a ground glass stopper on it, and cool it with a pump until it reaches room temperature. Number average molecular weight 300 g mol -1 The monomer polyethylene glycol methacrylate (PEGMA) (4.0 g, 13.340 mmol) was dissolved in ethanol / water (95 / 5 v / v; 8.75 mL), added to the vial, and the initiator α-bromo Methyl propionate (50 mu L, 0.440 mmol), ligand 2,2′-bipyridine (138.9 mg, 0.880 mmol), cuprous chloride (44.0 mg, 0.440 mmol) with a purity of 99.99% were added to the above mixed system, and the high-purity Argon protection. Raise the temperature to 30°C, and maintain this temperature, stir with magnetic force, and react for about 10-12 hours.

[0092] The reaction product was diluted by adding absolute ethanol, and contacted with a...

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Abstract

The invention discloses a bi-component gene carrier. In the invention, firstly, Fe3O4 (ferroferric oxide) magnetic nanoparticles which have magnetic response and are modified by cationic polymers are synthesized for condensing DNA (deoxyribonucleic acid); meanwhile, hydrophilic sulfydryl-containing brush type anion segmented copolymers are designed to be an external layer of magnetic cation carriers and coated on the external surface of the Fe3O4 magnetic nanoparticles modified by the cationic polymers to form an anion layer with an anti-non-specific adsorption performance; in addition, the sulfydryl is crosslinked to obtain disulfide bonds under the action of hydrogen peroxide or oxygen in airs, thus improving the stability of grains in the blood circulation process; and when the carrierenters the inside of cells, the disulfide bonds break in a cell reducing environment, thus the DNA can realize in-situ release in living cells. The bi-component gene carrier can be compounded with the DNA by electrostatic interaction to form a gene carrier with the magnetic response, the anti-protein absorption performance, the low toxicity and the transfection performance.

Description

technical field [0001] The invention belongs to the field of gene carriers and relates to a two-component gene carrier with magnetic responsiveness, protein adsorption resistance, low toxicity and transfection. Background technique [0002] In the prior art, it is often necessary to use molecular biology methods to introduce a target gene into target cells to express the target gene product. The key to this process is to select the appropriate gene carrier and gene introduction method, so that the target gene can be expressed safely, controllably, efficiently and stably in the target cells. [0003] Therefore, a gene carrier with stability, safety and targeting is firstly needed. The gene carriers currently used for targeted gene delivery include viruses, organic cationic complexes, recombinant proteins, and inorganic nanoparticles. Recombinant proteins have good biocompatibility in theory, but they are expensive to prepare and immature; inorganic nanoparticles are easy to ...

Claims

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Application Information

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IPC IPC(8): C12N15/87C08F120/34C08F293/00C08F8/34
Inventor 倪沛红郝莹张明祖朱秀林
Owner SUZHOU UNIV
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