Generation of a FLNA knockout hESC line (WAe009-A-P) to model cardiac valvular dysplasia using CRISPR/Cas9

Fengyuan Lu; Yifeng Gao; En Li

doi:10.1016/j.scr.2023.103162

Generation of a FLNA knockout hESC line (WAe009-A-P) to model cardiac valvular dysplasia using CRISPR/Cas9

Stem Cell Res. 2023 Sep:71:103162. doi: 10.1016/j.scr.2023.103162. Epub 2023 Jul 1.

Authors

Fengyuan Lu¹, Yifeng Gao¹, En Li²

Affiliations

¹ Department of Cardiology, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
² Department of Cardiology, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China. Electronic address: lidongfs@zzu.edu.cn.

PMID: 37429070
DOI: 10.1016/j.scr.2023.103162

Abstract

The FLNA gene encodes the cytoskeletal protein filamin A which plays a key role in the structure and function of the cardiac valves. Truncating FLNA mutations are associated with cardiac valvular dysplasia. To further understand the exact role of FLNA in this disease, we have generated a human FLNA knockout cell line from H9 using CRISPR/Cas9 technology in this study. This cell line WAe009-A-P has a 2 bp deletion in the exon 2 of FLNA gene which resulted in a frameshift in the translation of FLNA and no FLNA protein was detected in this cell line. Moreover, WAe009-A-P also expressed pluripotency markers, had a normal female karyotype (46XX) and maintained the ability to differentiate into the three germ layers in vitro.

Publication types

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

MeSH terms

CRISPR-Cas Systems* / genetics
Cell Line
Female
Heart Valves
Human Embryonic Stem Cells* / metabolism
Humans
Mutation