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A simple, quick, and efficient CRISPR/Cas9 genome editing method for human induced pluripotent stem cells.

Abstract：

:Induced pluripotent stem cells (iPSCs) have become an essential research platform to study different human diseases once being discovered by Dr. Shinya Yamanaka in 2006. Another breakthrough in biomedical research is the application of CRISPR/Cas9 system for genome editing in mammalian cells. Although numerous studies have been done to develop methods for gene editing in iPSCs, the current approaches suffer from several limitations, including time and labor consuming, low editing efficiency, and potential off-target effects. In the current study, we report an electroporation-mediated plasmid CRISPR/Cas9 delivery approach for genome editing in iPSCs. With this approach, an edited iPSC cell line could be obtained within 2 weeks. In addition, the transit introducing of CRISPR/Cas9 machinery could minimize genomic integration of Cas9 gene, which avoided potential long-term side effects of Cas9 enzyme. We showed that CRISPR/Cas9-mediated genomic editing did not affect pluripotency and differentiation ability of iPSCs. With the quickly evolving of both iPSC and CRISPR/Cas9-mediated genome editing research fields, we believe that our method can significantly facilitate the application of genome editing in iPSCs research.

journal_name

Acta Pharmacol Sin

journal_title

Acta pharmacologica Sinica

authors

Geng BC,Choi KH,Wang SZ,Chen P,Pan XD,Dong NG,Ko JK,Zhu H

doi

10.1038/s41401-020-0452-0

subject

Has Abstract

pub_date

2020-11-01 00:00:00

pages

1427-1432

issue

11

eissn

1671-4083

issn

1745-7254

pii

10.1038/s41401-020-0452-0

journal_volume

41

pub_type

杂志文章

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