Comprehensive Protocols for CRISPR/Cas9-based Gene Editing in Human Pluripotent Stem Cells.
Accepted version
Peer-reviewed
Repository URI
Repository DOI
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Authors
Santos, DP
Kiskinis, E
Eggan, K
Merkle, FT
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. © 2016 by John Wiley & Sons, Inc.
Description
Keywords
StemCellInstitute, CRISPR, gene editing, knock-in, pluripotent, stem cell
Journal Title
Curr Protoc Stem Cell Biol
Conference Name
Journal ISSN
1941-7322
1938-8969
1938-8969
Volume Title
38
Publisher
Wiley
Publisher DOI
Sponsorship
Medical Research Council (MC_PC_12009)
MRC (MR/P501967/1)
Academy of Medical Sciences (SBF001\1016)
National Institute of Neurological Disorders and Stroke (K99NS083713)
MRC (MR/P501967/1)
Academy of Medical Sciences (SBF001\1016)
National Institute of Neurological Disorders and Stroke (K99NS083713)
National Institutes of Health (HL109525, 5P01GM099117, 5K99NS083713), Wellcome Trust, Academy of Medical Sciences, Medical Research Council (MR/P501967/1), Les Turner ALS Foundation, Target ALS, Muscular Dystrophy Association