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Protocol for scarless genome editing of human pluripotent stem cell based on orthogonal selective reporters.
Zhao, Yun; Pan, Zihang; Hong, Zixuan; Sun, Mengze; Hong, Yi; Peng, Xiaohong; Li, Xiaoxia; Wang, Xi; Wang, Kai.
Affiliation
  • Zhao Y; Department of Physiology and Pathophysiology, School of Basic Medical Sciences, State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing 100191, China. Electronic address: zy157@pku.edu.cn.
  • Pan Z; Department of Physiology and Pathophysiology, School of Basic Medical Sciences, State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing 100191, China.
  • Hong Z; Department of Physiology and Pathophysiology, School of Basic Medical Sciences, State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing 100191, China.
  • Sun M; Department of Physiology and Pathophysiology, School of Basic Medical Sciences, State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing 100191, China.
  • Hong Y; Department of Physiology and Pathophysiology, School of Basic Medical Sciences, State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing 100191, China.
  • Peng X; Department of Physiology and Pathophysiology, School of Basic Medical Sciences, State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing 100191, China.
  • Li X; Department of Physiology and Pathophysiology, School of Basic Medical Sciences, State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing 100191, China.
  • Wang X; State Key Laboratory of Female Fertility Promotion, Clinical Stem Cell Research Center, Peking University Third Hospital, Beijing 100191, China. Electronic address: xi_wang@bjmu.edu.cn.
  • Wang K; Department of Physiology and Pathophysiology, School of Basic Medical Sciences, State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing 100191, China. Electronic address: kai.wang88@pku.edu.cn.
STAR Protoc ; 5(2): 103084, 2024 Jun 21.
Article in En | MEDLINE | ID: mdl-38787727
ABSTRACT
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.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Pluripotent Stem Cells / Gene Editing Limits: Humans Language: En Journal: STAR Protoc Year: 2024 Document type: Article Country of publication:
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Pluripotent Stem Cells / Gene Editing Limits: Humans Language: En Journal: STAR Protoc Year: 2024 Document type: Article Country of publication: