Generation of an EFNB2-2A-mCherry reporter human embryonic stem cell line using CRISPR/Cas9-mediated site-specific homologous recombination

被引：0

作者：

Huang, Ying ^{[1
,2
]}

Wu, Hongchun ^{[1
,2
]}

Han, Xinglong ^{[1
,2
]}

Wu, Jie ^{[1
,2
]}

Yu, Miao ^{[1
,2
]}

Zhao, Zhen-Ao ^{[3
,4
,5
]}

Shen, Zhenya ^{[1
,2
]}

Hu, Shijun ^{[1
,2
]}

Lei, Wei ^{[1
,2
]}

机构：

[1] Soochow Univ, Affiliated Hosp 1, Dept Cardiovasc Surg, Suzhou 215000, Peoples R China

[2] Soochow Univ, Med Coll, Inst Cardiovasc Sci, Collaborat Innovat Ctr Hematol,State Key Lab Radi, Suzhou 215000, Peoples R China

[3] Hebei North Univ, Inst Microcirculat, Zhangjiakou 075000, Peoples R China

[4] Hebei North Univ, Dept Pathophysiol, Basic Med Coll, Zhangjiakou 075000, Peoples R China

[5] Hebei Key Lab Crit Dis Mech & Intervent, Zhangjiakou 075000, Peoples R China

来源：

STEM CELL RESEARCH | 2021年 / 52卷

基金：

国家重点研发计划; 中国国家自然科学基金;

关键词：

D O I：

10.1016/j.scr.2021.102241

中图分类号：

Q813 [细胞工程];

学科分类号：

摘要：

Ephrin B2 (EFNB2) is the first identified and most widely used marker for arterial endothelial cells (AECs). We generated a heterozygous EFNB2-2A-mCherry reporter H1 cell line, H1-EFNB2-2A-mCherry(+/-)(WAe001-A-57), by CRISPR/Cas9-mediated insertion of 2A-mCherry cassette into the EFNB2 gene locus, immediately before the translation stop codon. The H1-EFNB2-2A-mCherry reporter cells were pluripotent and could differentiate into all three germ layer lineages. Simultaneous expression of mCherry was observed when expression of EFNB2 was increased during endothelial cell differentiation. Thus, the generated reporter cells enable live identification of EFNB2-positive AECs, and screening of small molecule compound and target genes that promote AEC differentiation.

引用

页数：5

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