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

利用CRISPR/Cas9编辑技术构建GLI3复合杂合敲除人胚胎干细胞系WAe001-A-20

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

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.

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