CRISPR/Cas9 technology mediated down producing goat VEGF gene fixed-point knock-in method

A cashmere goat, gene technology, applied in the field of animal molecular breeding, can solve the problem of lack of integrity
CN108570479AActive Publication Date: 2018-09-25INNER MONGOLIA UNIVERSITY
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
INNER MONGOLIA UNIVERSITY
Publication Date
2018-09-25

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Abstract

The invention provides a method for finishing down producing goat VEGF gene fixed-point knock-in mediated by a CRISPR / Cas9 system. The method comprises the following steps: establishing a gRNA expression vector and a VEGF homologous recombination vector on the basis of the CRISPR / Cas9 system according to a CCR5 gene sequence of down producing goats; then jointly transferring three optimized vectors into fibroblasts of down producing goat fetuses to obtain VEGF gene fixed-point knocked-in positive cells; and preparing VEGF gene fixed-point integrated down producing goats by using a somatic nuclear transfer technology. The established targeting vector based on the CRISPR / Cas9 system provides a simple, rapid and safe path for fixed-point knock-in of goat VEGF genes. The method does not involve any screening marker genes in a cell line screening process, therefore, the safety of transgenic animals is greatly improved, and the method has high value on research of genetic breeding and gene functions of down producing goats.
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Description

technical field

[0001] The invention belongs to the field of animal molecular breeding, in particular to a method for obtaining a transgenic cashmere goat by using CRISPR / Cas9 technology to integrate VEGF gene at a specific point. Background technique

[0002] In recent biological research, genome editing (Genome editing) technology is the latest research technology for researchers to understand the function of specific genes. With the completion of whole genome sequencing of more and more species, the new challenge facing researchers is how to obtain gene-related function and application information from a large amount of data. Gene editing technology is a powerful research tool to accomplish this important goal, for which it was named one of the top ten important scientific advances by Science in 2012. In recent years, genome-directed editing has developed rapidly. The production of artificial endonuclease (engineered endonuclease, EEN) has changed the limitations in the...

Claims

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