Preparation of CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 nano gene system and application of CRISPR/Cas9 nano gene system in transfection

Abstract

The invention belongs to the technical field of biology and particularly relates to preparation and cell transfection application of a CS-based (chitosan-based) and CRISPR / Cas9 plasmid-constructed nano gene drug. The preparation comprises dissolving chitosan into acetic acid solution, dropwise adding the chitosan acetic acid solution into water, adding in CRISPR in mass proportion to the chitosan,encapsulating and carrying CRISPR / Cas9 with the chitosan to prepare a novel gene drug nano particle for good gene transfection effects or targeted exogenous gene introduction and further to achieve the effects of transferring gene drugs into cells. The novel nano compound of CRISPR@CS structured on the CS and the CRIPPR through an optimal feeding ratio can significantly improve the transfection efficiency of genes in cells to convey more genes into the cells to take effects, thereby being expected to being widely applied in the field of gene treatment through gene drug delivery.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the preparation and cell transfection application of a nano-gene drug constructed on the basis of chitosan (CS) and CRISPR / Cas9 plasmid. Background technique [0002] Chitin exists in a very high amount in nature and is rich in sources. It is a biological polysaccharide second only to cellulose on the earth. It is widely found in animals, plants, and various fungi. Studies have found that especially crustaceans (crabs, shrimps, etc.) ) content is the highest, up to 58%~85%, followed by Insecta, etc., whose content can reach 20%~60%; Also contains a certain amount of chitin. Chitosan is a derivative obtained after the N-acetylamino group on the sugar group in chitin is removed. Generally speaking, chitosan is a straight chain connected by β-(1,4) glycosidic bonds. For each unit, more than 55% of the N-acetylamino group can be called chitosan, or in other words, the amoun...

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide a preparation of a novel CRISPR / Cas9 nano-gene system and its application in transfection, so as to solve the difficulty in quality control of traditional transfection vectors in the prior art and the inconspicuous transfection effect and low efficiency. The problem

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