利用CRISPR/Cas9编辑技术构建ESRG Pr-tdTomato报告基因人胚胎干细胞系CSUe011-A。
Generation of an ESRG Pr-tdTomato reporter human embryonic stem cell line, CSUe011-A, using CRISPR/Cas9 editing.
作者信息
Liu Hui, Li Shasha, Ren Caiping, Liu Weidong, Zhu Bin, Wang Lei, Xu Hongjuan, Xie Wen, Zuo Xiang, Zhou Yao, Luo Longlong, Jiang Xingjun
机构信息
Cancer Research Institute, Department of Neurosurgery, School of Basic Medical Science, Xiangya Hospital, Central South University, Changsha, China; Laboratory of Stem Cell & Retinal Regeneration, Institute of Stem Cell Research, Division of Ophthalmic Genetics, The Eye Hospital, Wenzhou Medical University, Wenzhou, China; The Key Laboratory of Carcinogenesis of the Chinese Ministry of Health and the Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Central South University, Changsha, China.
Cancer Research Institute, Department of Neurosurgery, School of Basic Medical Science, Xiangya Hospital, Central South University, Changsha, China; The Key Laboratory of Carcinogenesis of the Chinese Ministry of Health and the Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Central South University, Changsha, China.
出版信息
Stem Cell Res. 2020 Oct;48:101983. doi: 10.1016/j.scr.2020.101983. Epub 2020 Sep 3.
ESRG was first identified in our previous study. It is highly expressed in human embryonic stem cells (hESCs), whereas it is significantly down-regulated in differentiated cells and is undetectable in adult tissues. To develop an hESC line for monitoring the expression of ESRG for further study of its function, we used gene editing techniques to insert fusion sequences of ESRG promoter and tdTomato fluorescent protein gene into the AAVS1 human safe harbor locus. The gene-edited line had a normal karyotype, expressed pluripotency markers and differentiation potential.
ESRG最初是在我们之前的研究中被鉴定出来的。它在人类胚胎干细胞（hESCs）中高度表达，而在分化细胞中显著下调，在成人组织中无法检测到。为了开发一种用于监测ESRG表达以进一步研究其功能的hESC系，我们使用基因编辑技术将ESRG启动子和tdTomato荧光蛋白基因的融合序列插入到AAVS1人类安全港位点。基因编辑后的细胞系具有正常的核型，表达多能性标志物并具有分化潜能。
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