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当前位置: 首页 > 文章 > TALEN介导的CXCR4敲除肝癌细胞株的建立 生物技术通报 2016,32 (2) 225-228
Position: Home > Articles > The Establishment of HepG2 Cell Line with TALEN-mediated Knockout of CXCR4 Biotechnology Bulletin 2016,32 (2) 225-228

TALEN介导的CXCR4敲除肝癌细胞株的建立

作  者：
张文美;丁妍;郭兴荣;李东升;赵万红;王小莉
单  位：
湖北医药学院生物医学工程学院;湖北医药学院附属太和医院胚胎干细胞研究湖北省重点实验室
关键词：
TALEN;CXCR4;肝癌;基因打靶
摘  要：
通过TALEN打靶建立CXCR4的细胞株,旨在研究CXCR4对肝癌的影响。选用肝癌细胞株Hep G2,采用转录激活样效应物核酸酶(TALEN)干扰细胞中CXCR4的表达。构建的CXCR4 TALEN质粒转入Hep G2细胞,通过T7E1酶切确定打靶效率为40%,并通过测序筛选出了CXCR4敲除的单克隆细胞,免疫荧光和Western blot进一步证实CXCR4基因表达显著下调。
译  名：
The Establishment of HepG2 Cell Line with TALEN-mediated Knockout of CXCR4
作  者：
ZHANG Wen-mei;DING Yan;GUO Xing-rong;LI Dong-sheng;ZHAO Wan-hong;WANG Xiao-li;Hubei Key Laboratory of Embryonic Stem Cell Research,Hubei University of Medicine;College of Biomedical Engineering,Hubei University of Medicine;
关键词：
TALEN;;CXCR4;;hepatocellular carcinoma cell;;gene target
摘  要：
It was to research on the effect of CXCR4 on Hep G2 cells, we established one cell line which could down-regulate CXCR4 expression stably by transcription activaor-like effector nuclease(TALEN) gene targeting. The hepatocellular carcinoma cell line Hep G2 was chosen for this study, and the expression of CXCR4 was interfered by transcription activator-like effector nuclease(TALEN). The constructed CXCR4 TALEN plasmids were transfected into Hep G2 cells, and the targeting efficiency was determined as 40% by enzyme digestion of T7E1. The single clone cell of CXCR4 knockout was screened by gene sequencing, and the significant decrease of gene CXCR4 expression was confirmed by immunofluorescence and Western blot. The hepatocellular carcinoma cell line with stably down-regulating CXCR4 expression was established successfully by TALEN gene targeting.

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