Efficient Gene Editing of Human Induced Pluripotent Stem Cells Using CRISPR/Cas9

  • Saniye Yumlu
  • Sanum Bashir
  • Jürgen Stumm
  • Ralf KühnEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1961)


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.

Key words

Pluripotent stem cells Gene editing CRISPR Cas9 Knockout Knockin i53 Trex2 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Saniye Yumlu
    • 1
    • 2
  • Sanum Bashir
    • 1
    • 2
  • Jürgen Stumm
    • 1
    • 2
  • Ralf Kühn
    • 1
    • 2
    Email author
  1. 1.Max-Delbrück-Centrum für Molekulare MedizinBerlinGermany
  2. 2.Berlin Institute of HealthBerlinGermany

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