Genome Editing in Human Pluripotent Stem Cells: Approaches, Pitfalls, and Solutions

Cell Stem Cell. 2016 Jan 7;18(1):53-65. doi: 10.1016/j.stem.2015.12.002.


Human pluripotent stem cells (hPSCs) with knockout or mutant alleles can be generated using custom-engineered nucleases. Transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 nucleases are the most commonly employed technologies for editing hPSC genomes. In this Protocol Review, we provide a brief overview of custom-engineered nucleases in the context of gene editing in hPSCs with a focus on the application of TALENs and CRISPR/Cas9. We will highlight the advantages and disadvantages of each method and discuss theoretical and technical considerations for experimental design.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Alleles
  • CRISPR-Cas Systems*
  • DNA / analysis
  • Deoxyribonucleases / genetics
  • Deoxyribonucleases / metabolism
  • Genetic Techniques*
  • Genetic Vectors
  • Genome, Human*
  • Genotype
  • Humans
  • Mutation
  • Pluripotent Stem Cells / cytology*
  • Point Mutation
  • Sequence Homology, Nucleic Acid


  • DNA
  • Deoxyribonucleases