利用CRISPR/Cas9技术生成KCNJ11纯合敲除人类胚胎干细胞系WAe001-A-12
Generation of a KCNJ11 homozygous knockout human embryonic stem cell line WAe001-A-12 using CRISPR/Cas9.
作者信息
Yuan Fang, Guo Dongsheng, Gao Ge, Liu Yanli, Xu Yingying, Wu Yuhang, Yang Fan, Ke Xinrong, Lai Keyu, Hong Liangqing, Li Yin-Xiong
机构信息
School of Life Sciences, University of Science and Technology of China, Hefei, China; Institute of Public Health, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China; Key Laboratory of Regenerative Biology, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China; Guangdong Provincial Key Laboratory of Biocomputing, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.
Institute of Public Health, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China; Key Laboratory of Regenerative Biology, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China; Guangdong Provincial Key Laboratory of Biocomputing, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China; University of Chinese Academy of Sciences, Beijing, China.
出版信息
Stem Cell Res. 2017 Oct;24:89-93. doi: 10.1016/j.scr.2017.08.016. Epub 2017 Aug 30.
The ATP-sensitive potassium channel is an octameric complex, and one of its subunits, namely Kir6.2, is encoded by the KCNJ11 gene. Mutations in KCNJ11 result in hyperinsulinism or diabetes mellitus, associated with abnormal insulin secretion. Here, using CRISPR/Cas9 editing, we established a homozygous mutant KCNJ11 cell line, WAe001-A-12, which was generated by a 62-bp deletion in the coding sequence of the human embryonic stem cell line H1. It was confirmed that this deletion in the KCNJ11 gene did not affect the protein expression levels of key pluripotent factors. Additionally, normal karyotype and differentiation potency were observed for the cell line.
ATP敏感性钾通道是一种八聚体复合物，其亚基之一，即Kir6.2，由KCNJ11基因编码。KCNJ11突变会导致高胰岛素血症或糖尿病，与胰岛素分泌异常有关。在此，我们利用CRISPR/Cas9编辑技术建立了一个纯合突变KCNJ11细胞系WAe001-A-12，该细胞系是通过人胚胎干细胞系H1编码序列中62个碱基对的缺失产生的。已证实KCNJ11基因中的这种缺失不影响关键多能因子的蛋白质表达水平。此外，该细胞系具有正常的核型和分化能力。
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