System for efficient screening of candidate sgRNAs for CRISPR/CAS9 gene editing system from plants and application
A technology of CAS9 and testing system, applied in the direction of DNA/RNA fragments, genetic engineering, recombinant DNA technology, etc., can solve the problem of inability to directly reflect the real cleavage of sgRNA target sites, low cleavage efficiency of sgRNA target sites, and inability to achieve high Throughput screening and other issues, to achieve the effect of convenient construction method, low cost and high detection throughput
Inactive Publication Date: 2017-08-11
江苏贝瑞利生物科技有限公司
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The invention discloses a system for high-throughput screening of candidate sgRNA target sites for a CRISPR/CAS9 system and application thereof. The testing system comprises 1, plasmids for expressing multiple candidate sgRNAs at the same time; and 2, plasmids for expressing CAS9. Fast high-throughput identification of the cutting efficiency of the candidate sgRNA target sites can be achieved by means of an arabidopsis and rice protoplast transient expression system. It is important to select a target sequence capable of achieving efficient cutting before the CRISPR/CAS9 system knocks out or edits genes, the cutting efficiency of multiple candidate sgRNA target sites can be verified within a short period of time, so that the optimal sgRNA can be selected to improve the success rate of knockout, in this way, the working cost can be reduced, and working efficiency can be improved.
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