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

Yifang Xie; Daqi Wang; Feng Lan; Gang Wei; Ting Ni; Renjie Chai; Dong Liu; Shijun Hu; Mingqing Li; Dajin Li; Hongyan Wang; Yongming Wang

doi:10.1038/s41598-017-02456-y

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

Yifang Xie
, Daqi Wang
, Feng Lan
, Gang Wei
, Ting Ni
, Renjie Chai
, Dong Liu
, Shijun Hu
, Mingqing Li
, Dajin Li
, Hongyan Wang^*
, Yongming Wang

^*此作品的通讯作者

Fudan University
Capital Medical University
Southeast University, Nanjing
Nantong University
Soochow University
Children's Hospital of Fudan University and Shanghai Key Laboratory of Medical Epigenetics

科研成果: 期刊稿件 › 文章 › 同行评审

107 引用（Scopus）

摘要

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.

源语言	英语
期刊论文编号	2320
期刊	Scientific Reports
卷	7
期	1
DOI	https://doi.org/10.1038/s41598-017-02456-y
出版状态	已出版 - 1 12月 2017
已对外发布	是

联合国可持续发展目标

此成果有助于实现下列可持续发展目标：

可持续发展目标 3 良好健康与福祉

访问文件

10.1038/s41598-017-02456-y

其它文件与链接

链接到 Scopus 的出版物

引用此

@article{4072de45e2f740ee8a1251f1d7dd0759,

title = "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.",

author = "Yifang Xie and Daqi Wang and Feng Lan and Gang Wei and Ting Ni and Renjie Chai and Dong Liu and Shijun Hu and Mingqing Li and Dajin Li and Hongyan Wang and Yongming Wang",

note = "Publisher Copyright: {\textcopyright} 2017 The Author(s).",

year = "2017",

month = dec,

day = "1",

doi = "10.1038/s41598-017-02456-y",

language = "English",

volume = "7",

journal = "Scientific Reports",

issn = "2045-2322",

publisher = "Nature Research",

number = "1",

}

TY - JOUR

T1 - An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells

AU - Xie, Yifang

AU - Wang, Daqi

AU - Lan, Feng

AU - Wei, Gang

AU - Ni, Ting

AU - Chai, Renjie

AU - Liu, Dong

AU - Hu, Shijun

AU - Li, Mingqing

AU - Li, Dajin

AU - Wang, Hongyan

AU - Wang, Yongming

PY - 2017/12/1

Y1 - 2017/12/1

N2 - 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.

AB - 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.

UR - https://www.scopus.com/pages/publications/85019885327

U2 - 10.1038/s41598-017-02456-y

DO - 10.1038/s41598-017-02456-y

M3 - Article

C2 - 28539611

AN - SCOPUS:85019885327

SN - 2045-2322

VL - 7

JO - Scientific Reports

JF - Scientific Reports

IS - 1

M1 - 2320

ER -

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

摘要

联合国可持续发展目标

访问文件

其它文件与链接

学术指纹

引用此