@article{3e7bcad49ded41e2834c9581862b689f,
title = "Comprehensive protocols for CRISPR/Cas9-based gene editing in human pluripotent stem cells",
abstract = "Genome editing of human pluripotent stem cells (hPSCs) with the CRISPR/Cas9 system has the potential to revolutionize hPSC-based disease modeling, drug screening, and transplantation therapy. Here, we aim to provide a single resource to enable groups, even those with limited experience with hPSC culture or the CRISPR/Cas9 system, to successfully perform genome editing. The methods are presented in detail and are supported by a theoretical framework to allow for the incorporation of inevitable improvements in the rapidly evolving gene-editing field. We describe protocols to generate hPSC lines with gene-specific knock-outs, small targeted mutations, or knock-in reporters.",
keywords = "CRISPR, Gene editing, Knock-in, Pluripotent, Stem cell",
author = "Santos, {David P.} and Evangelos Kiskinis and Kevin Eggan and Merkle, {Florian T.}",
note = "Funding Information: We thank Feng Zheng, Jack Sandoe, Lindy Barrett, James Gagnon, Summer Thyme, Belinda von Niederh{\"a}usern, and Werner Neuhausser for their assistance with developing methodologies for CRISPR/Cas9-based gene editing. Peter Kirwan, Ralitsa Mad-sen, and Nuno Rocha provided useful comments that improved this manuscript. K.E.and F.T.M. were supported by grants from the National Institutes of Health (HL109525, 5P01GM099117, 5K99NS083713). F.T.M. is supported by funds from the Wellcome Trust, the Academy of Medical Sciences, and the Medical Research Council (MR/P501967/1). E.K. and D.S. are supported by grants from the Les Turner ALS Foundation, Target ALS, and the Muscular Dystrophy Association. Publisher Copyright: {\textcopyright} 2016 by John Wiley & Sons, Inc.",
year = "2016",
month = aug,
doi = "10.1002/cpsc.15",
language = "English (US)",
volume = "2016",
journal = "Current Protocols in Stem Cell Biology",
issn = "1941-7322",
publisher = "John Wiley and Sons Inc.",
}
