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携带多西环素诱导型Cas9表达盒的人扩展多能干细胞系hEPS1-iCas9-B建立

Generation of a human extended pluripotent stem cell line (SKLRMe002-A) carrying a doxycycline-inducible Cas9 expression cassette

Zhazheng He, Ke Feng, Hao Sun, Tianyi Yu, Dicong Zhu, Yang Yang

Journal:Stem Cell Research

IF:1.59

DOI:10.1016/j.scr.2022.102816

PMID:35617843

Published:2022-05-17

research field:分子生物学转化医学肿瘤免疫学免疫治疗RNA生物学

Abstract

Human extended pluripotent stem cell (hEPS) is a novel type of pluripotent stem cell, which possesses bi-potency towards both embryonic and extraembryonic lineages. Here, we generated a hEPS cell line (hEPS1-iCas9-B) from the cell line named hEPS1, carrying a doxycycline-inducible Cas9 expression cassette along with a constitutive reverse tetracycline transactivator (M2rtTA) expression cassette at the AAVS1 locus, thus we could efficiently generate genetically modified hEPS for studies. This cell lined remained self-renewal, differentiation potential and normal karyotype. Meanwhile, it showed robust transcriptional expression of Cas9 with doxycycline induction and could target the site where the sgRNA guided.

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