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通过CRISPR/Cas9编辑构建GLI3复合杂合敲除人类胚胎干细胞系WAe001-A-20

Construction of a GLI3 compound heterozygous knockout human embryonic stem cell line WAe001-A-20 by CRISPR/Cas9 editing.

作者信息

Wei Ruzhi, Yuan Fang, Wu Yuhang, Liu Yanli, You Kai, Yang Zhen, Chen Yan, Getachew Anteneh, Wang Ning, Xu Yingying, Zhuang Yuanqi, Yang Fan, Li Yin-Xiong

机构信息

Institute of Public Health, 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; 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; University of Chinese Academy of Sciences, Beijing, China.

Institute of Public Health, 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; 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.

出版信息

Stem Cell Res. 2018 Oct;32:139-144. doi: 10.1016/j.scr.2018.09.010. Epub 2018 Sep 17.

Abstract

The human GLI3 protein has a dual function as a transcriptional activator or repressor of hedgehog signaling, depending on the proteolytic processing forms of GLI3. In this study, we established a compound heterozygous GLI3 mutant human embryonic stem cell line (WAe001-A-20) through CRISPR/Cas9 editing. The WAe001-A-20 cells carried two deletions on two different alleles of exon 2 of GLI3, respectively, which resulted in a frame shift and early termination in the translation of GLI3. Moreover, WAe001-A-20 maintains a normal karyotype, parental cell morphology, pluripotent phenotype and the ability to differentiate into three germ layers. Resource table.

摘要

人类GLI3蛋白具有双重功能,根据GLI3的蛋白水解加工形式,它可以作为刺猬信号通路的转录激活因子或抑制因子。在本研究中,我们通过CRISPR/Cas9编辑建立了一种复合杂合GLI3突变人类胚胎干细胞系(WAe001-A-20)。WAe001-A-20细胞在GLI3外显子2的两个不同等位基因上分别有两个缺失,这导致了GLI3翻译中的移码和早期终止。此外,WAe001-A-20保持正常的核型、亲代细胞形态、多能表型以及分化为三个胚层的能力。资源表。

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