Comprehensive protocols for CRISPR/Cas9-based gene editing in human pluripotent stem cells

David P. Santos, Evangelos Kiskinis, Kevin Eggan, Florian T. Merkle*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

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.

Original languageEnglish (US)
JournalCurrent Protocols in Stem Cell Biology
Volume2016
DOIs
StatePublished - Aug 2016

Keywords

  • CRISPR
  • Gene editing
  • Knock-in
  • Pluripotent
  • Stem cell

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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