Construction method of Mia3 conditional gene knockout mouse model

Abstract

The invention relates to a method for constructing a Mia3 gene conditional knockout mouse model by using a CRISPR (clustered regularly interspaced short palindromic repeats) / Cas9 (CRISPR associated protein 9) technology. The method comprises the following steps: 1, designing sgRNA (small guide ribonucleic acid) aiming at mouse Mia3 gene Exon2-Exon7, 2, obtaining the sgRNA by using an in-vitro transcription technology, and 3, co-injecting or co-electrically transferring a targeting vector, the sgRNA and Cas9 protein to a mouse fertilized egg. Compared with a traditional ES targeting mouse model, the Mia3 gene conditional knockout mouse model constructed for the first time has the characteristics of high efficiency, rapidness, simplicity, convenience, low cost and the like, and the expression of the gene in a specific tissue or specific time can be regulated through Cre.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a method for constructing a Mia3 conditional gene knockout mouse model and an application thereof. Background technique [0002] CRISPR / Cas9 technology achieves specific DNA recognition by designing specific sgRNA, and completes cutting at the targeted position, and then completes the repair of the break through the cell's own DNA repair mechanism, thereby realizing the "editing" of the target gene. With the development of CRISPR / Cas9 technology and the deepening of research, it has been widely used to construct transgenic model animals (mainly gene-edited mice), so as to provide basic research on animal-level treatment before clinical trials. [0003] Both the research on the pathogenesis of human diseases and the screening of effective therapeutic drugs require a large number of preclinical trials. Due to the ethical constraints of directly using human cells and ...

AI Technical Summary

Problems solved by technology

[0007] At present, there are no relevant reports on the construction of Mia3 gene conditional knockout mice using CRISPR / Cas9 technology

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