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Construction method for Sqstm1 whole-genome knockout mouse animal model and application

A gene knockout mouse, animal model technology, applied in the biological field to achieve the effect of economic animal model

Inactive Publication Date: 2018-11-06
SHANGHAI TONGJI HOSPITAL
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However, there is no report on the construction method of the Sqstm1 knockout mouse animal model

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[0044] The present invention will be further described below in combination with specific embodiments. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, it should be understood that after reading the contents of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

[0045] 1. This implementation category involves a method for establishing a Sqstm1 gene knockout mouse model based on CRISPR / Cas9 gene knockout technology. The technical route is as follows: figure 1 shown.

[0046] 2. Determine the basic information of the knockout gene

[0047] 1. Knockout gene name (MGI number): Sqstm1 (MGI: 107931)

[0048]2. Knockout gene MGI website link: http: / / www.informatics.jax.org / marker / MGI...

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Abstract

The invention discloses a construction method for a Sqstm1 whole-genome knockout mouse animal model and application. The mouse animal model is a mouse of which the Sqstm1 gene is knocked out. On the basis of a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) / Cas9 gene knockout technology, the Sqstm1 gene knockout mouse model is constructed. The construction method comprises the following steps: S1: designing sgRNA (Ribonucleic Acid) and Cas9 RNA, carrying out in vitro transcription to obtain mRNA, carrying out microinjection on the sgRNA and the Cas9 RNA with activity into amouse oosperm, to obtain the Sqtm1 gene knockout mouse; S2: identifying the Sqtm1 gene knockout mouse animal model. On the basis of the CRISPR / Cas9 gene knockout technology, the Sqstm1 gene knockout mouse animal model is constructed for the first time, and a convenient, reliable and economic animal model is provided for researching a relationship between Sqstm1 and diseases including autophagy, tumors and the like.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a construction method and application of a Sqstm1 whole gene knockout mouse animal model. Background technique [0002] Sqstm1 gene, full name Sequestosome 1, also known as p62; A170; DMRV gene, etc., the gene is located on chromosome 5 q35, the molecular weight of Sqstm1 is 62kD, and it exists in the cytoplasm in the form of scattered dots or aggregates. It consists of 440 amino acids Coded, including PBl domain, TB domain, UBA domain, LIR domain and other functional domains. The existence of these functional domains makes Sqstm1 the core of multiple signaling pathways, and the abnormal function of Sqstm1 will lead to the blockage of multiple signaling pathways, abnormal protein aggregation and induce the occurrence of various diseases. [0003] Sqstm1 was once identified as an independent ligand for tyrosine phosphorylation. With the deepening of research, more and more evidences ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/90A01K67/027
CPCC12N15/113A01K67/0276A01K2207/15A01K2217/075A01K2227/105A01K2267/0331C07K14/47C12N15/907C12N2310/10C12N2310/20
Inventor 李冬吴军录孙祖俊权文强
Owner SHANGHAI TONGJI HOSPITAL
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