利用CRISPR/Cas9技术构建CYP2E1基因敲除的HEK293FT细胞系

doi:10.3969/j.issn.1004-616x.2017.05.006

Abstract

Abstract: OBJECTIVE: To construct the CYP2E1 knock-out (KO) HEK293FT cell line using CRISPR/Cas9 system and explore its usefulness in chemical toxicity testing. METHODS: The sgRNA expression vector targeting CYP2E1 gene was transfected into HEK293FT cells. Single cells from the transfected cells were isolated through serial dilutions to get the HEK293FT CYP2E1 KO cell line. The cell line was tested for expression of cytotoxicity from N-(4-hydroxyphenyl)-acetamide(4-APAP) and 1,2-dichloroethane(1,2-DCE). RESULTS: Two HEK293FT CYP2E1 KO cell lines were obtained and they showed dramatic reduction in susceptibility to N-(4-hydroxyphenyl)-acetamide (4-APAP) and 1,2-dichloroethane (1,2-DCE) cytotoxicity. CONCLUSION: HEK293FT CYP2E1 KO cell lines were successfully constructed using a CRISPR/Cas9 system and they can be useful for studying the function of CYP2E1.

Key words: CYP2E1, CRISPR/Cas9, gene knockout, HEK293FT cell line

CLC Number:

R392.11

FAN Qiming, LI Ruobi, WANG Fei, GUO Tao, WANG Ting, LI Daochuan, XING Xiumei, CHEN Liping, CHEN Wen, WANG Qing. Construction of CRISPR/Cas9-mediated genomic deletion of the CYP2E1 gene in human HEK293FT cell line[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(5): 352-357,363.

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Construction of CRISPR/Cas9-mediated genomic deletion of the CYP2E1 gene in human HEK293FT cell line

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