Application of difference agency technique to C. T base substitution cell enrichment

A base, biological cell technology, applied in the application field of differential surrogate technology in the enrichment of C·T base replacement cells
CN110564752AActive Publication Date: 2019-12-13BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
Publication Date
2019-12-13

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Abstract

The invention discloses an application of a difference agency technique to C. T base substitution cell enrichment. A difference agency technique carrier disclosed by the invention comprises the following reagents: sgRNA, a C. T base substitution system and a functional incapacitation screening agent resistant gene, wherein the sgRNA consists of esgRNA targeting a target point sequence of an objective gene and sgRNA targeting a target point sequence of the functional incapacitation screening agent resistant gene; the C.T base substitution system can perform C. T base substitution on the targetpoint sequence of the functional incapacitation screening agent resistant gene under guidance of the sgRNA for targeting the target point sequence of the functional incapacitation screening agent resistant gene so that the functions of the functional incapacitation screening agent resistant gene are recovered. According to the application disclosed by the invention, C.T base substitution cell enrichment on cell level can be realized, and the C.T base substitution efficiency is greatly improved.
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Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to the application of differential agent technology in C·T base substitution cell enrichment. Background technique

[0002] CRISPR-Cas9 technology has become a powerful genome editing method and has been widely used in many tissues and cells. The CRISPR / Cas9 protein-RNA complex is positioned on the target by the guide RNA (guide RNA), cuts and generates a DNA double-strand break (dsDNA break, DSB), and then the organism will instinctively initiate a DNA repair mechanism to repair the DSB. There are generally two repair mechanisms, one is non-homologous end joining (NHEJ), and the other is homologous recombination (homology-directed repair, HDR). Usually NHEJ accounts for the majority, so the random indels (insertions or deletions) generated by the repair are much higher than the precise repair. For precise base substitution, the application of HDR to achieve precise base substitution ...

Claims

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