Vector for carrying out site-specific mutagenesis on MSTN (Myostatin) and site-specific integration on PPARgamma (Peroxisome Proliferator-activated Receptor gamma) at same time

A MRF-PPAR, site-directed mutagenesis technology, applied in vector, nucleic acid vector, DNA preparation, etc., can solve problems such as skeletal muscle increase, MSTN inactivity, destruction of three-dimensional structure stability and function, etc., to improve accuracy and efficiency, improve transgenic Efficiency, effect of shortening breeding time
CN109679998AActive Publication Date: 2019-04-26NORTHWEST A & F UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
NORTHWEST A & F UNIV
Publication Date
2019-04-26

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Abstract

The invention discloses a vector for carrying out site-specific mutagenesis on MSTN (Myostatin) and site-specific integration on PPARgamma (Peroxisome Proliferator-activated Receptor gamma) at the same time, and recombinant cells. The vector comprises a Cas9 cutting expression vector and a targeting vector. According to the vector disclosed by the invention, a recombinant cell strain can be constructed by carrying out co-transfection on host cells by the Cas9 cutting expression vector and the targeting vector, the recombinant cell strain can be used as nuclear transfer donor cells for producing transgenic cloned embryo, function exertion of the MSTN can be inhibited, and meanwhile, specific expression of the PPARgamma in skeletal muscle cells can be realized, so that support is provided for quickly and efficiently developing high-quality transgenic beef new products.
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Description

technical field

[0001] The present invention relates to the technical field of transgenic cloning animals, and more specifically relates to a vector and a recombinant cell for targeting site-directed mutation of MSTN gene through miRNA and simultaneously site-specific integration of PPARγ gene. Background technique

[0002] Myostatin (MSTN) is also called growth differentiation factor (GDF8). As a negative regulator of muscle growth and differentiation, it can inhibit the transcriptional activity of the positive regulator - myogenic determinant factor (MoyD). Inhibits proliferation and differentiation of myoblasts. Mature MSTN inhibited the DNA and protein synthesis of C2C12 myogenic cells cultured in vitro in a dose-sensitive manner, and could inhibit the mitotic transition from G1 phase to S phase and from G2 phase to M phase of C2C12 myogenic cells, as well as induce C2C12 myogenic cells. The autophagy of myogenic cells thus inhibits the proliferation of C2C12 myogenic c...

Claims

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