Generation of a homozygous P4HA2 knockout human embryonic stem cell line (FDCHDPe012-A) by CRISPR/Cas9 system

Jie You; Shuangping Ma; Haokun Zhang; Ling Chen

Stem Cell Research (Oct 2022)

Generation of a homozygous P4HA2 knockout human embryonic stem cell line (FDCHDPe012-A) by CRISPR/Cas9 system

Jie You,
Shuangping Ma,
Haokun Zhang,
Ling Chen

Affiliations

Jie You: Department of Ophthalmology & Vision Science, Eye & ENT Hospital, Shanghai Medical School, Fudan University, Shanghai, China; Key NHC Key Laboratory of Myopia (Fudan University), Laboratory of Myopia, Chinese Academy of Medical Sciences, China; Shanghai Key Laboratory of Visual Impairment and Restoration, Eye & ENT Hospital, Shanghai Medical School, Fudan University, Shanghai, China
Shuangping Ma: Institutes of Health Central Plains, Xinxiang Medical University, Xinxiang, Henan, China
Haokun Zhang: State Key Laboratory of Genetic Engineering, MOE Engineering Research Center of Gene Technology, School of Life Sciences, Fudan University, Shanghai 200438, China
Ling Chen: Department of Ophthalmology & Vision Science, Eye & ENT Hospital, Shanghai Medical School, Fudan University, Shanghai, China; Key NHC Key Laboratory of Myopia (Fudan University), Laboratory of Myopia, Chinese Academy of Medical Sciences, China; Shanghai Key Laboratory of Visual Impairment and Restoration, Eye & ENT Hospital, Shanghai Medical School, Fudan University, Shanghai, China; Corresponding author at: Department of Ophthalmology & Vision Science, Eye & ENT Hospital, Fudan University, Shanghai, China.

Journal volume & issue: Vol. 64
p. 102930

Abstract

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Prolyl 4-hydroxylase subunit alpha-2(P4HA2) is associated with autosomal dominant high myopia. A significant reduction of P4HA2 protein expression has been observed in fibroblast cells of high myopia patients with inherited P4HA2 mutations. To explore the function of P4HA2 in cases of high myopia, we generated a P4HA2-KO hES cell line (FDCHDPe012-A) efficiently through CRISPR/Cas9 system. We confirmed the presence of a 5-bp biallelic deletion, causing a frameshift and premature P4HA2 translation termination. The FDCHDPe012-A presented morphology of typical stem cells expressed pluripotent genes, possessed a normal parental karyotype, and could differentiate into three germ layers.

Published in Stem Cell Research

ISSN: 1873-5061 (Print); 1876-7753 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Science: Biology (General)
Website: https://www.journals.elsevier.com/stem-cell-research

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