title: An iCRISPR platform for rapid, multiplexable, and inducible genome editing in human pluripotent stem cells.
abstract: Human pluripotent stem cells (hPSCs) offer a unique platform for elucidating the genes and molecular pathways that underlie complex traits and diseases. To realize this promise, methods for rapid and controllable genetic manipulations are urgently needed. By combining two newly developed gene-editing tools, the TALEN and CRISPR/Cas systems, we have developed a genome-engineering platform in hPSCs, which we named iCRISPR. iCRISPR enabled rapid and highly efficient generation of biallelic knockout hPSCs for loss-of-function studies, as well as homozygous knockin hPSCs with specific nucleotide alterations for precise modeling of disease conditions. We further demonstrate efficient one-step generation of double- and triple-gene knockout hPSC lines, as well as stage-specific inducible gene knockout during hPSC differentiation. Thus the iCRISPR platform is uniquely suited for dissection of complex genetic interactions and pleiotropic gene functions in human disease studies and has the potential to support high-throughput genetic analysis in hPSCs.
authors.affiliation: Developmental Biology Program, Sloan-Kettering Institute, 1275 York Avenue, New York, New York 10065, USA.
authors.forename: Federico
authors.initials: F
authors.lastname: González
authors.affiliation: Developmental Biology Program, Sloan-Kettering Institute, 1275 York Avenue, New York, New York 10065, USA.
authors.forename: Zengrong
authors.initials: Z
authors.lastname: Zhu
authors.affiliation: Developmental Biology Program, Sloan-Kettering Institute, 1275 York Avenue, New York, New York 10065, USA.
authors.forename: Zhong-Dong
authors.initials: ZD
authors.lastname: Shi
authors.affiliation: Developmental Biology Program, Sloan-Kettering Institute, 1275 York Avenue, New York, New York 10065, USA.
authors.forename: Katherine
authors.initials: K
authors.lastname: Lelli
authors.affiliation: Developmental Biology Program, Sloan-Kettering Institute, 1275 York Avenue, New York, New York 10065, USA.|Weill Graduate School of Medical Sciences at Cornell University/The Rockefeller University/Sloan-Kettering Institute Tri-Institutional M.D.-Ph.D. Program, 1300 York Avenue, New York, NY 10065, USA.
authors.forename: Nipun
authors.initials: N
authors.lastname: Verma
authors.affiliation: Developmental Biology Program, Sloan-Kettering Institute, 1275 York Avenue, New York, New York 10065, USA.|Louis V. Gerstner Jr. Graduate School of Biomedical Sciences, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10065, USA.
authors.forename: Qing V
authors.initials: QV
authors.lastname: Li
authors.affiliation: Developmental Biology Program, Sloan-Kettering Institute, 1275 York Avenue, New York, New York 10065, USA.
authors.forename: Danwei
authors.initials: D
authors.lastname: Huangfu
mesh_terms: D000483:Alleles; D001483:Base Sequence; D002454:Cell Differentiation; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004277:DNA, Single-Stranded / Q000235:genetics; D055786:Gene Knockout Techniques; D018390:Gene Targeting; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D016678:Genome; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014162:Transfection
chemical_list: D004277:DNA, Single-Stranded