Home > Browse Issues > Vol.36 No.12
Generation of HtrA2 Knockout Cell Line with An Improved CRISPR/Cas9 System
Hu Yuan, Zhang Ting, Jiang Chang’an*
State Key Laboratory of Biotherapy/Collaborative Innovation Center of Biotherapy, Sichuan University, Chengdu 610041, China
Abstract: Sequence-specific nucleases are powerful tools for genome editing. The RNA-guided Cas9 nuclease from Streptococcus pyogenes can be effectively targeted to genomic loci and generate DNA breaks. The site-specific DNA cleavages of Cas9 rely on the core 20 nt targeting sequences within its guide RNA (sgRNA or gRNA). In this paper， we describe a modified version of the CRISPR/Cas9 system， which significantly simplifies the construction of gRNA expression vectors. Using this system， we have efficiently generated HtrA2 knockout HEK293T cells by removing the first exon of the gene. This improvement significantly reduces the cost associated with the genome editing using CRISPR/CAS9 system.
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