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

Journal: 
Nat Methods
Publication Year: 
2014
Authors: 
Yuichiro Miyaoka
Amanda H Chan
Luke M Judge
Jennie Yoo
Miller Huang
Trieu D Nguyen
Paweena P Lizarraga
Po-Lin So
Bruce R Conklin
PubMed link: 
24509632
Public Summary: 
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.
Scientific 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.