CRISPR/Cas9 technology based preparation method and applications of PI3K[gamma] whole body knockout mouse and kit

Abstract

The invention provides a CRISPR / Cas9 technology based preparation method and applications of a PI3K[gamma] whole body knockout mouse and a kit. The preparation method comprises following steps: designing sgRNA for identify the gene PI3K[gamma]; connecting the sgRNA to a plasmid carrier, carrying out in-vitro transcription to obtain RNA that can be applied to microinjection; injecting the RNA intoa mouse fertilized egg, after injection, giving birth to mice; identifying the genotype of the mice, selecting mice, whose PI3K[gamma] whole body knockout genotype detection result is positive, and mating picked mice with wild mice to obtain PI3K[gamma] whole body knockout mice with a stable genotype. Compared with a conventional gene knockout technology, the operation is easier, the efficiency ishigher, homozygote mutation is obtained more easily, and good foundation is provided for the research on functions of PI3K[gamma].

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for preparing PI3Kγ whole-body knockout model mice based on CRISPR / Cas9 technology, an application thereof and a kit. Background technique [0002] Gene knockout is aimed at a sequence whose sequence is known but whose function is unknown, changing the genetic gene of the organism, making the specific gene function ineffective, so that part of the function is blocked, and can further affect the organism, and then it is speculated that the The biological functions of genes. [0003] Gene knockout technology was developed in the 1980s. It is a new molecular biology technology based on gene homologous recombination technology and embryonic stem cell technology. Gene homologous recombination technology refers to the homologous recombination of exogenous DNA and genes with the same or similar sequence in the genome of the recipient cell, thereby replacing the same / similar gene seq...

AI Technical Summary

Problems solved by technology

[0007] In order to overcome at least one deficiency of the prior art, the present invention provides a method for preparing PI3Kγ whole-body knockout model mice based on CRISPR / Cas9 technology, using RNA-guided Cas9 nuclease to cut PI3Kγ at the PI3Kγ genomic site of mouse fertilized eggs Gene, and then obtain homozygous PI3Kγ knockout mice through procedures such as embryo transfer and mouse crossing, in order to overcome the disadvantages of low production efficiency, long production cycle and high failure rate of PI3Kγ knockout mice

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