Targeted Gene Editing in Human Pluripotent Stem Cells Using Site-Specific Nucleases

  • Sylvia Merkert
  • Ulrich Martin
Part of the Advances in Biochemical Engineering/Biotechnology book series (ABE, volume 163)


Introduction of induced pluripotent stem cell (iPSC) technology and site-directed nucleases brought a major breakthrough in the development of regenerative therapies and biomedical research. With the advancement of ZFNs, TALENs, and the CRISPR/Cas9 technology, straightforward and precise manipulation of the genome of human pluripotent stem cells (PSC) became possible, allowing relatively easy and fast generation of gene knockouts, integration of transgenes, or even introduction of single nucleotide changes for correction or introduction of disease-specific mutations. We review current applications of site-specific nucleases in human PSCs and focus on trends and challenges for efficient gene editing and improvement of targeting strategies.

Graphical Abstract


CRISPR/Cas9 Homologous recombination NHEJ TALEN ZFN 



Self-cleaving peptide sequence


Adeno-associated virus integration site 1 (safe harbor site)


ATP-binding cassette subfamily A member 1


AKT serine/threonine protein kinase 2


Amyotrophic lateral sclerosis




Cytomegalovirus early enhancer element coupled to chicken beta-actin promoter


CRISPR-associated system


C-C motif chemokine receptor 5


Citrate lyase beta-like


Clustered regularly interspaced short palindromic repeats


DNA methyltransferase 3B


Double-strand break


Enhancer of zeste homolog 2


GATA binding protein 4


His 3, Lys 4 or Lys 9


Homologous recombination


Immunodeficiency-centromeric region instability-facial anomalies syndrome


Induced pluripotent stem cell


Myosin heavy chain


Neurogenein 3


Nonhomologous end joining


Octamer-binding protein 4


Protospacer adjacent motif


Pluripotent stem cells


Sortilin 1


Single-stranded oligonucleotide


Transcription activator-like effector nuclease


Zinc-finger nuclease



We thank R. Zweigerdt for contributing to fruitful discussions and critical reading of the manuscript, and R. Olmer for providing microscopy images of endothelial cells.


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Copyright information

© Springer International Publishing AG 2017

Authors and Affiliations

  1. 1.Department of Cardiothoracic, Transplantation and Vascular SurgeryLeibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO)HannoverGermany
  2. 2.REBIRTH-Cluster of Excellence, German Center for Lung Research (DZL)GießenGermany
  3. 3.Hannover Medical SchoolHannoverGermany

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