Generation of a TPM1 homozygous knockout embryonic stem cell line by CRISPR/Cas9 editing

Shuhong Ma; Qi Xu; Rui Bai; Tao Dong; Zhiping Peng; Xujie Liu

doi:10.1016/j.scr.2021.102470

Generation of a TPM1 homozygous knockout embryonic stem cell line by CRISPR/Cas9 editing

Stem Cell Res. 2021 Aug:55:102470. doi: 10.1016/j.scr.2021.102470. Epub 2021 Jul 19.

Authors

Shuhong Ma¹, Qi Xu², Rui Bai¹, Tao Dong¹, Zhiping Peng³, Xujie Liu⁴

Affiliations

¹ Anzhen Hospital, Capital Medical University, Beijing 100029, China; Beijing Institute of Heart, Lung and Blood Vessel Diseases, Beijing 100029, China.
² State Key Laboratory of Cardiovascular Disease, National Center for Cardiovascular Diseases, Fuwai Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100037, China.
³ Department of Radiology, Basic Medical School, Chongqing Medical University, Chongqing 400016, China.
⁴ Department of Radiology, Basic Medical School, Chongqing Medical University, Chongqing 400016, China. Electronic address: xujie.liu@childrens.harvard.edu.

PMID: 34352617
DOI: 10.1016/j.scr.2021.102470

Abstract

Alpha-Tropomyosin (TPM1) plays a crucial role in actin regulation and stability and contributes fundamental functions to heart development: without TPM1 expressing, mice embryos will die early in embryogenesis. To further identify the role of TPM1 in human cardiac development, here we generated a homozygous TPM1 knockout (TPM1^-/-) human embryonic stem cell (hESC) line using CRISPR/Cas9-based genome editing system. The generated TPM1^-/- hESC line maintained normal karyotype, highly expressed pluripotency markers and was able to differentiate into all three germ layers in vivo. This cell line provides a powerful tool to investigate the role of TPM1 in heart development in future.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CRISPR-Cas Systems* / genetics
Cell Line
Embryonic Stem Cells
Human Embryonic Stem Cells*
Humans
Mice
Tropomyosin / genetics

Substances

Tropomyosin