Method for constructing high-quality porcine nuclear transplantation donor cells with high lean meat percentage, fast growth and resistance to porcine reproductive and respiratory syndrome and series diarrhea diseases and application of high-quality porcine nuclear transplantation donor cells

A technology for expressing vectors and vector skeletons, applied in biochemical equipment and methods, chemical instruments and methods, cells modified by introducing foreign genetic materials, etc., can solve the problem of inactivation of receptor proteins, inability to infect live pigs, and reduced infectivity And other issues
CN112779292AActive Publication Date: 2021-05-11NANJING KGENE GENETIC ENG CO LTD
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
NANJING KGENE GENETIC ENG CO LTD
Publication Date
2021-05-11

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Abstract

The invention discloses a method for constructing high-quality porcine nuclear transplantation donor cells with high lean meat percentage, fast growth and resistant to porcine reproductive and respiratory syndrome and series diarrhea diseases and application of the high-quality porcine nuclear transplantation donor cells. A CRISPR / Cas9 (clustered regularly interspaced short palindromic repeats / CRISPR associated protein 9) system for porcine MSTN-SST-CD163-pAPN four-gene editing contains a Cas9 expression vector and gRNA (guide Ribonucleic Acid) expression vectors aiming at a porcine MSTN gene, an SST gene, a CD163 gene and a pAPN gene, and plasmid complete sequence of the Cas9 expression vector is as shown in SEQ ID NO.2. According to the invention, the corresponding gRNA expression vectors are designed for different targets of MSTN, SST, CD163 and pAPN genes, and gRNA with relatively high editing efficiency and the gRNA expression vector are obtained through screening. Through cooperation with the modified Cas9 efficient expression vector for gene editing, the editing efficiency is remarkably improved as compared with that of an original vector.
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Description

technical field

[0001] The invention belongs to the field of biotechnology, and relates to a method for constructing high-quality pig nuclear transplantation donor cells with high lean meat rate, fast growth and resistance to blue ear disease and serial diarrhea diseases and its application, in particular to the use for MSTN, SST, CD163, pAPN Four-gene edited CRISPR / Cas9 system and its application in the construction of high-quality pig nuclear transfer donor cells with high lean meat percentage, fast growth and resistance to PRRS and serial diarrheal diseases. Background technique

[0002] Pig is one of the earliest domesticated domestic animals in my country, and it has always been an important meat animal for human beings in the long river of history. The Chinese love to eat pork is related to the food culture for thousands of years. Since 2000, pork has accounted for more than 70% of my country's meat consumption, and it is the most important meat consumed in my country...

Claims

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