Generation of a PDK-1 knockout human embryonic stem cell line by CRISPR/(WAe009-A-2K) Cas9 editing

Amina Saleem; Mingyu Wei; Muhammad Khawar Abbas; Siyao Zhang; Jiaqi Fan; Yang Xian; Hongfeng Jiang

doi:10.1016/j.scr.2024.103642

Generation of a PDK-1 knockout human embryonic stem cell line by CRISPR/(WAe009-A-2K) Cas9 editing

Stem Cell Res. 2024 Dec 28:83:103642. doi: 10.1016/j.scr.2024.103642. Online ahead of print.

Authors

Amina Saleem¹, Mingyu Wei², Muhammad Khawar Abbas³, Siyao Zhang⁴, Jiaqi Fan⁴, Yang Xian⁴, Hongfeng Jiang⁵

Affiliations

¹ Anzhen Hospital, Capital Medical University, Beijing 100029, China; Beijing Institute of Heart, Lung, and Blood Vessel Diseases, Beijing 100029, China.
² Department of Cardiology, Peking University Third Hospital, 49 Huayuan North Road, Haidian District, Beijing 100191, China.
³ Department of Human Anatomy, Histology and Embryology, School of Basic Medical Sciences, Capital Medical University, Beijing, China; Department of Homoeopathic Medical Sciences, Faculty of Medicine and Allied Health Sciences, The Islamia University of Bahawalpur, 63100 Punjab, Pakistan.
⁴ Anzhen Hospital, Capital Medical University, Beijing 100029, China; Maternal-Fetal Medicine Centre in Fetal Heart Disease, Beijing Anzhen Hospital, Capital Medical University, Beijing 100029, China.
⁵ Anzhen Hospital, Capital Medical University, Beijing 100029, China; Beijing Institute of Heart, Lung, and Blood Vessel Diseases, Beijing 100029, China. Electronic address: [email protected].

PMID: 39798272
DOI: 10.1016/j.scr.2024.103642

Abstract

Pyruvate Dehydrogenase Kinase1 (PDK1) belongs to the family of kinases, regulates diverse metabolic processes. PDK1 is a susceptibility locus for heart failure via thinning of ventricle walls, and enlarged atria and ventricles. We successfully developed a PDK1 knockout (PDK1^-/^-) human embryonic stem cell (hESC) line using an episomal vector-based CRISPR/Cas9 system explore the role of PDK in human heart development. This PDK1-KO hESC line-maintained stem cell-like morphology, pluripotency, and normal karyotype and can differentiate into all three germ layers in vivo. This cell line will be a valuable tool for future research on the role of PDK1 in heart development.