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Efficient gene editing of human induced pluripotent stem cells using CRISPR/Cas9

Authors

  • S. Yumlu
  • S. Bashir
  • J. Stumm
  • R. Kühn

Journal

  • Methods in Molecular Biology

Citation

  • Methods Mol Biol 1961: 137-151

Abstract

  • The generation of targeted mutants is a crucial step toward studying the biomedical effect of genes of interest. The generation of such mutants in human induced pluripotent stem cells (iPSCs) is of an utmost importance as these cells carry the potential to be differentiated into any cell lineage. Using the CRISPR/Cas9 nuclease system for induction of targeted double-strand breaks, gene editing of target loci in iPSCs can be achieved with high efficiency. This chapter covers protocols for the preparation of reagents to target loci of interest, the transfection, and for the genotyping of single cell-derived iPSC clones. Furthermore, we provide a protocol for the convenient generation of plasmids enabling multiplex gene targeting.


DOI

doi:10.1007/978-1-4939-9170-9_10