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 ve...

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Autores principales:	Yifang Xie, Daqi Wang, Feng Lan, Gang Wei, Ting Ni, Renjie Chai, Dong Liu, Shijun Hu, Mingqing Li, Dajin Li, Hongyan Wang, Yongming Wang
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/791c11cd5d1445dbb9c3d7cae88ee914
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spelling	oai:doaj.org-article:791c11cd5d1445dbb9c3d7cae88ee9142021-12-02T15:05:05ZAn episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells10.1038/s41598-017-02456-y2045-2322https://doaj.org/article/791c11cd5d1445dbb9c3d7cae88ee9142017-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-02456-yhttps://doaj.org/toc/2045-2322Abstract 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.Yifang XieDaqi WangFeng LanGang WeiTing NiRenjie ChaiDong LiuShijun HuMingqing LiDajin LiHongyan WangYongming WangNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yifang Xie Daqi Wang Feng Lan Gang Wei Ting Ni Renjie Chai Dong Liu Shijun Hu Mingqing Li Dajin Li Hongyan Wang Yongming Wang An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells
description	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.
format	article
author	Yifang Xie Daqi Wang Feng Lan Gang Wei Ting Ni Renjie Chai Dong Liu Shijun Hu Mingqing Li Dajin Li Hongyan Wang Yongming Wang
author_facet	Yifang Xie Daqi Wang Feng Lan Gang Wei Ting Ni Renjie Chai Dong Liu Shijun Hu Mingqing Li Dajin Li Hongyan Wang Yongming Wang
author_sort	Yifang Xie
title	An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells
title_short	An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells
title_full	An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells
title_fullStr	An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells
title_full_unstemmed	An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells
title_sort	episomal vector-based crispr/cas9 system for highly efficient gene knockout in human pluripotent stem cells
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/791c11cd5d1445dbb9c3d7cae88ee914
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An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells

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