通过 CRISPR/Cas9 介导的同源重组生成 RYBP FLAG-HA 敲入人胚胎干细胞系。
Generation of RYBP FLAG-HA knock-in human embryonic stem cell line through CRISPR/Cas9-mediated homologous recombination.
机构信息
CAS Key Laboratory of Regenerative Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China; University of Chinese Academy of Sciences, Beijing 100049, China; Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China; Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China.
CAS Key Laboratory of Regenerative Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China; Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
出版信息
Stem Cell Res. 2022 Jul;62:102803. doi: 10.1016/j.scr.2022.102803. Epub 2022 May 4.
RYBP, a critical component of polycomb repressive complex1 (PRC1), is required for the pluripotency and differentiation of mouse embryonic stem cells(mESCs). However, its function and mechanism to regulate human embryonic stem cells(hESCs) remain unknown. Here, to investigate the role of RYBP in hESCs, we generate an hESC line with FLAG-HA tag knock-in to RYBP locus through CRISPR/Cas9-mediated homologous recombination. hESC with RYBP_FLAG-HA knock-in maintains normal morphology and karyotype, while it maintains pluripotency to differentiate into three germ layers.
RYBP 是多梳抑制复合物 1（PRC1）的关键组成部分，对于小鼠胚胎干细胞（mESCs）的多能性和分化是必需的。然而，其调节人类胚胎干细胞（hESCs）的功能和机制尚不清楚。在这里，为了研究 RYBP 在 hESCs 中的作用，我们通过 CRISPR/Cas9 介导的同源重组在 RYBP 基因座产生了带有 FLAG-HA 标签敲入的 hESC 系。带有 RYBP_FLAG-HA 敲入的 hESC 保持正常的形态和核型，同时保持多能性并分化为三个胚层。
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