癌变·畸变·突变2017，Vol.29Issue(5)：352-357,363,7.DOI:10.3969/j.issn.1004-616x.2017.05.006
利用CRISPR/Cas9技术构建CYP2E1基因敲除的HEK293FT细胞系
Construction of CRISPR/Cas9-mediated genomic deletion of the CYP2E1 gene in human HEK293FT cell line
范启明 1王庆 1李若碧 1王飞 1郭涛 1王婷 1李道传 1邢秀梅 1陈丽萍 1陈雯1
作者信息
- 1. 中山大学公共卫生学院,广东 广州 510080
- 折叠
摘要
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.关键词
CYP2E1/CRISPR/Cas9/基因敲除/HEK293FT细胞系Key words
CYP2E1/CRISPR/Cas9/gene knockout/HEK293FT cell line分类
医药卫生引用本文复制引用
范启明,王庆,李若碧,王飞,郭涛,王婷,李道传,邢秀梅,陈丽萍,陈雯..利用CRISPR/Cas9技术构建CYP2E1基因敲除的HEK293FT细胞系[J].癌变·畸变·突变,2017,29(5):352-357,363,7.
