Construction method and application of Arvcf gene knockout animal model

Abstract

The invention relates to a method for constructing an Arvcf gene knockout mouse model and application. The mouse model is a mouse of which seven exons in an Arvcf gene are knocked out, and meanwhile, the invention discloses the method for constructing the Arvcf gene knockout mouse model. The method comprises the following steps of firstly, determining a knockout region of the gene Arvcf to be knocked out, then designing and synthesizing specific sgRNA and a corresponding primer, linearizing and purifying a constructed sgRNA/Cas9 vector, then carrying out in vitro transcription, finally injecting the sgRNA/Cas9 vector into fertilized eggs of the mouse through micro-injection, and transplanting the sgRNA/Cas9 vector into the uterus of a pseudopregnant female mouse to establish the Arvcf gene knockout mouse model. Tails of the obtained filial generation mouse are collected and subject to DNA extraction, and PCR reaction and sequencing are conducted twice by using two pairs of primers to confirm that the obtained mouse is a wild-type mouse or homozygous or heterozygous mouse of which Arvcf gene target regions are knocked out. The gene knockout mouse model can be used for researching the effect of the ARVCF in mental diseases in which the ARVCF is involved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a construction method and application of an Arvcf gene knockout animal model. Background technique [0002] The protein encoded by the ARVCF (Armadillo Repeat gene deleted in Velo-Cardio-Facial syndrome) gene is a member of the p120ctn catenin family, and other members include p120ctn, δ-catenin and p0071. This family of proteins plays a key role in the formation of adherens junction complexes and also plays an important role in various aspects of neuromorphogenesis, and members of this family have also been found to be associated with various neurodevelopmental diseases. In addition, the ARVCF protein is part of the cadherin-catenin complex, which may regulate cadherin-mediated junctional structure and cell-cell adhesion in various cell types, and the ARVCF gene has also been reported to be involved in nerves from ganglion eminences. Yuan migration. In addition, in the...

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Problems solved by technology

[0004] The purpose of the embodiment of the present invention is to provide a method and application for establishing an Arvcf gene knockout mouse model based on a new generation of gene editing technology, so as to solve the technical problems of the existing gene editing technology such as cumbersome operation and high cost, and at the same time, it can also solve the lack of Issues in Animal Models for Studying ARVCF Biological Function

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