Isolation of single-base genome-edited human iPS cells without antibiotic selection

Nat Methods. 2014 Mar;11(3):291-3. doi: 10.1038/nmeth.2840. Epub 2014 Feb 9.

Abstract

Precise editing of human genomes in pluripotent stem cells by homology-driven repair of targeted nuclease-induced cleavage has been hindered by the difficulty of isolating rare clones. We developed an efficient method to capture rare mutational events, enabling isolation of mutant lines with single-base substitutions without antibiotic selection. This method facilitates efficient induction or reversion of mutations associated with human disease in isogenic human induced pluripotent stem cells.

Publication types

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

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Base Composition / genetics
  • Cell Line
  • Cloning, Molecular
  • Cytological Techniques / methods*
  • Genome, Human*
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / drug effects
  • Mutation

Substances

  • Anti-Bacterial Agents