Fig. 1. Robust co-selection for prime editing.

a Schematic for the co-enrichment of CRISPR-driven editing events at a GOI. b Schematic representation of the ATP1A1 locus regions targeted by SpCas9. The first and third extracellular loops of the Na⁺/K⁺ ATPase are encoded by ATP1A1 exon 4 and 17, respectively. The relative levels of resistance to ouabain conferred by different ATP1A1 mutations in K562 cells are shown. c PE and small indels quantification as determined by BEAT and TIDE analysis from Sanger sequencing. K562 cells were transfected with PE3 vectors targeting ATP1A1 exon 17 (T804N) and the indicated GOI. Genomic DNA was harvested 3 days post-transfection (before selection) and cells were treated (ouabain) or not (untreated) with 0.5 µM ouabain until all non-resistant cells were eliminated. d Same as in c but co-targeting was performed via ATP1A1 exon 4 (Q118R). n = 2 independent biological replicates performed at different times. Source data are provided in the Source Data file.