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Curr Protoc Stem Cell Biol DOI:10.1002/cpsc.15

Comprehensive Protocols for CRISPR/Cas9-based Gene Editing in Human Pluripotent Stem Cells.

Publication TypeJournal Article
Year of Publication2016
AuthorsSantos, DP, Kiskinis, E, Eggan, K, Merkle, FT
JournalCurr Protoc Stem Cell Biol
Date Published2016 Aug 17
KeywordsCell Culture Techniques, Cell Line, Clone Cells, Cloning, Molecular, Colony-Forming Units Assay, CRISPR-Cas Systems, DNA, Gene Amplification, Gene Editing, Gene Knockout Techniques, Gene Targeting, Genes, Reporter, Genetic Vectors, Genome, Human, High-Throughput Nucleotide Sequencing, Humans, Mutation, Oligonucleotides, Plasmids, Pluripotent Stem Cells, Polymerase Chain Reaction, Reproducibility of Results, RNA, Guide, Transcription, Genetic

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.


Alternate JournalCurr Protoc Stem Cell Biol
PubMed ID27532820
PubMed Central IDPMC4988528
Grant ListP01 GM099117 / GM / NIGMS NIH HHS / United States
R01 HL109525 / HL / NHLBI NIH HHS / United States
/ / Wellcome Trust / United Kingdom
MC_PC_12009 / / Medical Research Council / United Kingdom
K99 NS083713 / NS / NINDS NIH HHS / United States