利用TALEN技术建立Mon1a基因敲除细胞系及其表型分析

Construction Mon1a Gene Knock-out HEK293T Stable Cell Line by TALEN and Phenotype Analysis

为了研究囊泡运输蛋白Mon1a(Mon1 secretory trafficking family member A)在细胞中的作用,该文利用类转录激活因子效应物核酸酶(transcription activator-like effector nuclease,TALEN)打靶技术构建了针对Mon1a基因的特定TALEN质粒对,转染后筛选获得Mon1a基因敲除的HEK293T稳定细胞系。进一步从细胞增殖能力、迁移能力以及小鼠皮下成瘤能力等方面研究了Mon1a缺失对HEK293T细胞的影响。结果表明,Mon1a在细胞增殖、迁移和成瘤等过程中起到重要作用。

In order to investigate the function of vesicular traffic protein Mon1a（Mon1 secretory trafficking family member A） in cells, we utilized TALEN（transcription activator-like effector nuclease） targeting technology to construct a pair of TALEN plasmids that aim at Mon1 a gene. After transfection, we successfully got the Mon1 agene knock-out HEK293 T stable cell line by screening. Then we detected the effects of Mon1 a in HEK293 T cell through the ability of proliferation, migration and the mice subcutaneous tumor formation by corresponding experiments. The results showed that loss of Mon1 a would inhibit cell proliferation, cell migration and tumor formation.