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Knockout TP53 of HeLa Cells by CRISPR/Cas9 Technology
Dong Wenjie1, Yang Yuanqin1, Fang Xianlong2, Yin Xiaofei2, Xu Yanni3, Wang Jiaojiao1,Chu Liang2, Wang Ruwei3, Fang Ling3, Wu Shuai2*, Liu Xinyuan1,2*
1Xinyuan Institute of Medicine and Biotechnology, Zhejiang Sci-Tech University, Hangzhou 310018, China;
2State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology,Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China;
3Zhejiang Conba Pharmaceutical Co., Ltd, Hangzhou 310018, China
2State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology,Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China;
3Zhejiang Conba Pharmaceutical Co., Ltd, Hangzhou 310018, China
Abstract: We generated TP53 knockout cells via CRISPR/Cas9 system in HeLa cells. CRISPR/Cas9 system could precisely knockout TP53 gene in HeLa cells and insert selection markers (as donor template for the recombination repairing) in DSBs (double-strand breaks) site as well. Further functional study revealed that HeLa (TP53–/–) cells get vigorous growth， resistance to chemotherapy and oxidative stress， implying that HeLa (TP53–/–) cells become more cancerous. All our data described a simple and efficient approach for gene deletion in cell line via the CRISPR/Cas9 system， largely helped clarifying the functions of gene and analyzing the mechanism of cellular process.
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