Fig. 4: Homozygous fluorescent DNA repair assay to visualize and quantify allele-specific DNA repair outcomes during biallelic editing in iPS cells.
From: Synergistic gene editing in human iPS cells via cell cycle and DNA repair modulation
a Schematic of the predicted outcomes from GFP editing in a homozygous AAVS1-CAG::EGFP (GFP) iPS cell line, and their distribution in FACS. b Comparison of monoallelic and biallelic editing in heterozygous and homozygous GFP iPS cells, showing DNA repair outcome frequency. HDR* indicates double-positive BFP/GFP cells arising from heterozygous editing and HDR repair in homozygous GFP iPS cells. c Ratio of HDR/MutEJ repair outcomes measured in b. d Quantification of biallelic repair outcomes in homozygous GFP iPS cells obtained from FACS-gating on each possible DNA repair outcome represented in a. e Effect of combined cold shock and drug treatment on repair outcome frequency in homozygous GFP iPS cells. f Ratio of HDR/MutEJ repair outcomes measured in e. g Biallelic repair outcomes in the absence (ā) or presence (Nā+āS) of combined drug treatment obtained from DNA repair outcomes represented in e. All data are presented as the meanāĀ±āS.D. of three technical replicates for each respective treatment. Source data are provided as a Source Data file.
