Protocol for scarless genome editing of human pluripotent stem cell based on orthogonal selective reporters

被引：1

作者：

Zhao, Yun ^{[1
]}

Pan, Zihang ^{[1
]}

Hong, Zixuan ^{[1
]}

Sun, Mengze ^{[1
]}

Hong, Yi ^{[1
]}

Peng, Xiaohong ^{[1
]}

Li, Xiaoxia ^{[1
]}

Wang, Xi ^{[2
]}

Wang, Kai ^{[1
]}

机构：

[1] Peking Univ, Sch Basic Med Sci, Dept Physiol & Pathophysiol, State Key Lab Vasc Homeostasis & Remodeling, Beijing 100191, Peoples R China

[2] Peking Univ Third Hosp, Clin Stem Cell Res Ctr, State Key Lab Female Fertil Promot, Beijing 100191, Peoples R China

来源：

STAR PROTOCOLS | 2024年 / 5卷 / 02期

基金：

北京市自然科学基金; 中国国家自然科学基金; 国家重点研发计划;

关键词：

GENERATION;

D O I：

10.1016/j.xpro.2024.103084

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Human pluripotent stem cells (hPSCs) hold great promise for applications in regenerative medicine and disease modeling. Here, we present a protocol for establishing edited hPSC cell lines utilizing visualized orthogonal selective reporters. We describe steps for constructing plasmids, carrying out cell culture and electroporation, as well as performing drug-fluorescent dual enrichment, clone screening, and cell line characterization. This protocol facilitates the achievement of single-base homozygous mutations and reporter knockins, offering a reliable approach for precision genome editing.

引用

页数：24

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