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An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells.

ABSTRACT: Human pluripotent stem cells (hPSCs) represent a unique opportunity for understanding the molecular mechanisms underlying complex traits and diseases. CRISPR/Cas9 is a powerful tool to introduce genetic mutations into the hPSCs for loss-of-function studies. Here, we developed an episomal vector-based CRISPR/Cas9 system, which we called epiCRISPR, for highly efficient gene knockout in hPSCs. The epiCRISPR system enables generation of up to 100% Insertion/Deletion (indel) rates. In addition, the epiCRISPR system enables efficient double-gene knockout and genomic deletion. To minimize off-target cleavage, we combined the episomal vector technology with double-nicking strategy and recent developed high fidelity Cas9. Thus the epiCRISPR system offers a highly efficient platform for genetic analysis in hPSCs.

SUBMITTER: Xie Y

PROVIDER: S-EPMC5443789 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature

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An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells.

Xie Yifang Y Wang Daqi D Lan Feng F Wei Gang G Ni Ting T Chai Renjie R Liu Dong D Hu Shijun S Li Mingqing M Li Dajin D Wang Hongyan H Wang Yongming Y

Scientific reports 20170524 1

Human pluripotent stem cells (hPSCs) represent a unique opportunity for understanding the molecular mechanisms underlying complex traits and diseases. CRISPR/Cas9 is a powerful tool to introduce genetic mutations into the hPSCs for loss-of-function studies. Here, we developed an episomal vector-based CRISPR/Cas9 system, which we called epiCRISPR, for highly efficient gene knockout in hPSCs. The epiCRISPR system enables generation of up to 100% Insertion/Deletion (indel) rates. In addition, the e ...[more]

PMID: 28539611

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