Gene editing method for plasmid vector group, dna group and foreign gene introduction

Abstract

The invention belongs to the technical field of genetic engineering and molecular precision improvement and breeding of poultry breeds, and discloses a gene editing method for introducing a plasmid vector group, a DNA group and an exogenous gene, including: P1 plasmid, P2 plasmid and P3 plasmid; DNA components include: The RNA-mediated Cas9 endogenous endonuclease gene, the specific site-mediated sgRNA gene and the site of introduction of the gene fragment and the insertion site-specific homologous DNA fragment. The method of the present invention can integrate the target DNA fragment into the appropriate position of the genome of the poultry organism, and it is helpful to insert the target DNA fragment into the appropriate position of the animal cell genome, thereby obtaining transgenic or chimeric animal individuals. The carrier group of the present invention has the characteristics of easy construction, easy mass production, safety and high efficiency.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering and molecular precision improvement and breeding of poultry varieties, and in particular relates to a gene editing method for introducing plasmid vector groups, DNA groups and exogenous genes. Background technique [0002] The egg industry is a major characteristic of my country's poultry industry, especially the egg duck industry. Therefore, there are many related breeding studies at home and abroad. However, in the production process of the egg industry, the proportion of male birds used is extremely low (the ratio of male to female in the case of artificial insemination). Only 1:20~30), so the hatching production of male birds during the hatching process is the biggest waste of resources in the development of the egg and poultry industry. Therefore, the question is how to use the latest gene editing technology to introduce foreign genes for the labeling of fertilized eggs of male bird...

AI Technical Summary

Problems solved by technology

[0003] To sum up, the problems existing in the existing technology are: due to the common problems in lentiviral transfection at present, the introduction of lentiviruses in traditional transgenic operations is random, and the position of introducing foreign genes is uncertain, because the introduction position causes Poultry mutation; lentivirus operation is very harmful and needs to be exposed to the virus during the operation process, which is harmful to the operator; the operation is too complicated

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