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.
基金：
National Natural Science Foundation of ChinaNational Natural Science Foundation of China [81370302, 81430005, 81270232, 31521003]; National Basic Research Program of ChinaNational Basic Research Program of China [2015CB943300]; Foundation for Innovative Research Group of the National Natural Science Foundation of ChinaNational Natural Science Foundation of China [31521003]; Open Research Funds of the State Key Laboratory of Genetic Engineering Fudan University [SKLGE-1407]; Ministry of Education of ChinaMinistry of Education, China [KPH1322054]; Thousand Youth Talents Plan Project of Thousand Youth Talents; 973 ProgramNational Basic Research Program of China [2013CB945403]; Commission for Science and Technology of Shanghai MunicipalityScience & Technology Commission of Shanghai Municipality (STCSM) [13JC1407600]
第一作者机构：[1]Fudan Univ, Shanghai Med Coll, Inst Biomed Sci, Shanghai 200032, Peoples R China;
通讯作者：
通讯机构：[2]Fudan Univ, State Key Lab Genet Engn, Sch Life Sci, Shanghai 200438, Peoples R China;[3]Fudan Univ, MOE Key Lab Contemporary Anthropol, Sch Life Sci, Shanghai 200438, Peoples R China;[4]Fudan Univ, Key Lab Reprod Regulat NPFPC SIPPR, Inst Reprod & Dev, Obstet & Gynecol Hosp, Shanghai 200011, Peoples R China;[10]Fudan Univ, Childrens Hosp, 399 Wanyuan Rd, Shanghai 201102, Peoples R China
推荐引用方式(GB/T 7714)：
Xie Yifang,Wang Daqi,Lan Feng,et al.An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells[J].SCIENTIFIC REPORTS.2017,7(1):-.doi:10.1038/s41598-017-02456-y.
APA：
Xie, Yifang,Wang, Daqi,Lan, Feng,Wei, Gang,Ni, Ting...&Wang, Yongming.(2017).An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells.SCIENTIFIC REPORTS,7,(1)
MLA：
Xie, Yifang,et al."An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells".SCIENTIFIC REPORTS 7..1(2017):-
