Generation of SHMT2 knockout human embryonic stem cell line (WAe009-A-67) using CRISPR/Cas9 technique

Jisheng Li; Bowen Zhang; Min Gan; Yunxing Li; Lijuan He; Wen Yue; Haixuan Qiao; Xuetao Pei; Yanhua Li

doi:10.1016/j.scr.2021.102581

Generation of SHMT2 knockout human embryonic stem cell line (WAe009-A-67) using CRISPR/Cas9 technique

Stem Cell Res. 2021 Dec:57:102581. doi: 10.1016/j.scr.2021.102581. Epub 2021 Oct 19.

Authors

Jisheng Li¹, Bowen Zhang², Min Gan¹, Yunxing Li³, Lijuan He⁴, Wen Yue⁴, Haixuan Qiao⁵, Xuetao Pei⁶, Yanhua Li⁷

Affiliations

¹ Stem Cell and Regenerative Medicine Lab, Institute of Health Service and Transfusion Medicine, Beijing 100850, China.
² Beijing Institute of Radiation Medicine, Beijing 100850, China; South China Research Center for Stem Cell & Regenerative Medicine, SCIB, Guangzhou 510005, China.
³ Beijing Institute of Radiation Medicine, Beijing 100850, China.
⁴ Stem Cell and Regenerative Medicine Lab, Institute of Health Service and Transfusion Medicine, Beijing 100850, China; South China Research Center for Stem Cell & Regenerative Medicine, SCIB, Guangzhou 510005, China.
⁵ School of Biomedical Engineering and Technology, Tianjin Medical University, Tianjin 300070, China. Electronic address: qiaohaixuan@aliyun.com.
⁶ Stem Cell and Regenerative Medicine Lab, Institute of Health Service and Transfusion Medicine, Beijing 100850, China; South China Research Center for Stem Cell & Regenerative Medicine, SCIB, Guangzhou 510005, China. Electronic address: peixt@bmi.ac.cn.
⁷ Beijing Institute of Radiation Medicine, Beijing 100850, China; South China Research Center for Stem Cell & Regenerative Medicine, SCIB, Guangzhou 510005, China. Electronic address: liyanhua@scrm.org.cn.

PMID: 34688993
DOI: 10.1016/j.scr.2021.102581

Abstract

Serine hydroxymethyltransferase 2 (SHMT2), a catalytic enzyme playing an important role in aerobic cellular respiration and mitochondrial metabolism, might be pivotal in self-renewal and differentiation of human pluripotent stem cells. Herein, we used the CRISPR/Cas9 editing system to construct a homozygous SHMT2 knockout (SHMT2-KO) human embryonic stem cell (hESC) line, exhibiting a normal karyotype, colony morphology, and high expression levels of pluripotent proteins. Furthermore, SHMT2 knockout did not impact the self-renewal ability or differentiation potential into three germ layers of hESCs. Accordingly, this cell line provides a valuable model for further assessing SHMT2 functions in human embryonic development.