title: Efficient Genome Editing in Induced Pluripotent Stem Cells with Engineered Nucleases In Vitro.
abstract: Precision genome engineering is rapidly advancing the application of the induced pluripotent stem cells (iPSCs) technology for in vitro disease modeling of cardiovascular diseases. Targeted genome editing using engineered nucleases is a powerful tool that allows for reverse genetics, genome engineering, and targeted transgene integration experiments to be performed in a precise and predictable manner. However, nuclease-mediated homologous recombination is an inefficient process. Herein, we describe the development of an optimized method combining site-specific nucleases and the piggyBac transposon system for "seamless" genome editing in pluripotent stem cells with high efficiency and fidelity in vitro.
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, 94305, USA.
authors.forename: Vittavat
authors.initials: V
authors.lastname: Termglinchan
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, 94305, USA.
authors.forename: Timon
authors.initials: T
authors.lastname: Seeger
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, 94305, USA.
authors.forename: Caressa
authors.initials: C
authors.lastname: Chen
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, 94305, USA.|Department of Medicine, Division of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, CA, 94305, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, 94305, USA.
authors.forename: Joseph C
authors.initials: JC
authors.lastname: Wu
authors.affiliation: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, 94305, USA. ioannis1@stanford.edu.|Department of Cardiothoracic Surgery, Stanford University School of Medicine, Stanford, CA, 94305, USA. ioannis1@stanford.edu.
authors.forename: Ioannis
authors.initials: I
authors.lastname: Karakikes
mesh_terms: D001483:Base Sequence; D064113:CRISPR-Cas Systems; D002469:Cell Separation; D002999:Clone Cells; D004720:Endonucleases / Q000378:metabolism*; D000072669:Gene Editing / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000378:metabolism; D057809:HEK293 Cells; D059765:Homologous Recombination; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D000069896:Transcription Activator-Like Effector Nucleases; D014162:Transfection
chemical_list: D004720:Endonucleases; D000069896:Transcription Activator-Like Effector Nucleases
keywords: CRISPR/Cas9; Genome engineering; Homology-directed repair; PiggyBac transposon system; TALEN