Generation of a Junctophilin-2 homozygous knockout human embryonic stem cell line (WAe009-A-36) by an episomal vector-based CRISPR/Cas9 system

Fujian Wu; Xiaowei Li; Rui Bai; Ya'nan Li; Jian Gao; Feng Lan

doi:10.1016/j.scr.2020.101930

Generation of a Junctophilin-2 homozygous knockout human embryonic stem cell line (WAe009-A-36) by an episomal vector-based CRISPR/Cas9 system

Stem Cell Res. 2020 Oct:48:101930. doi: 10.1016/j.scr.2020.101930. Epub 2020 Aug 3.

Authors

Fujian Wu¹, Xiaowei Li², Rui Bai¹, Ya'nan Li¹, Jian Gao³, Feng Lan⁴

Affiliations

¹ Beijing Laboratory for Cardiovascular Precision Medicine, MOE Key Laboratory of Medical Engineering for Cardiovascular Diseases, MOE Key Laboratory of Remodeling-Related Cardiovascular Disease, Beijing Collaborative Innovation Center for Cardiovascular Disorders, Anzhen Hospital, Capital Medical University, Beijing 100029, China; Beijing Institute of Heart, Lung and Blood Vessel Diseases, Beijing 100029, China.
² Department of Cardiology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
³ University of British Columbia, Vancouver, Canada.
⁴ Beijing Laboratory for Cardiovascular Precision Medicine, MOE Key Laboratory of Medical Engineering for Cardiovascular Diseases, MOE Key Laboratory of Remodeling-Related Cardiovascular Disease, Beijing Collaborative Innovation Center for Cardiovascular Disorders, Anzhen Hospital, Capital Medical University, Beijing 100029, China; Beijing Institute of Heart, Lung and Blood Vessel Diseases, Beijing 100029, China. Electronic address: fenglan@ccmu.edu.cn.

PMID: 32777767
DOI: 10.1016/j.scr.2020.101930

Abstract

Mutations in Junctophilin-2(JPH2) gene is the cause of hypertrophic cardiomyopathy (HCM) and leading inherited cause of left ventricular hypertrophy and myofilaments disarray. JPH2 protein, a member of the Junctophilin family, is mainly expressed in heart and plays an important role in E-C coupling. We have generated a homozygous JPH2 knockout (JPH2-KO) human embryonic stem cell (hESC) line using an episomal vector-based CRISPR/Cas9 system. This JPH2-KO hESC line maintained stem cell like morphology, pluripotency, normal karyotype and could differentiate into all three germ layers in vivo.

Publication types

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

MeSH terms

CRISPR-Cas Systems / genetics
Cell Line
Human Embryonic Stem Cells*
Humans
Membrane Proteins / genetics

Substances

Membrane Proteins
junctophilin

Grants and funding

MOP142265 /CIHR/Canada