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

被引：0
|
作者：
You, Jie [1 ,2 ,3 ]
Ma, Shuangping [4 ]
Zhang, Haokun [5 ]
Chen, Ling [1 ,2 ,3 ,6 ]
机构：
[1] Fudan Univ, Eye & ENT Hosp, Shanghai Med Sch, Dept Ophthalmol & Vis Sci, Shanghai, Peoples R China
[2] Fudan Univ, Chinese Acad Med Sci, Key NHC Key Lab Myopia, Lab Myopia, Shanghai, Peoples R China
[3] Fudan Univ, Eye & ENT Hosp, Shanghai Med Sch, Shanghai Key Lab Visual Impairment & Restorat, Shanghai, Peoples R China
[4] Xinxiang Med Univ, Inst Hlth Cent Plains, Xinxiang, Henan, Peoples R China
[5] Fudan Univ, MOE Engn Res Ctr Gene Technol, Sch Life Sci, State Key Lab Genet Engn, Shanghai 200438, Peoples R China
[6] Fudan Univ, Eye & ENT Hosp, Dept Ophthalmol & Vis Sci, Shanghai, Peoples R China
来源：
STEM CELL RESEARCH | 2022年 / 64卷
基金：
中国国家自然科学基金;
关键词：
D O I：
10.1016/j.scr.2022.102930
中图分类号：
Q813 [细胞工程];
学科分类号：
摘要：
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.
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页数：5
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