Supplementary Figure 4: Selection for CRISPR–Cas9-driven targeted mutagenesis by coediting ATP1A1 via NHEJ.
From: Marker-free coselection for CRISPR-driven genome editing in human cells
(a) Experimental strategy for the co-enrichment of CRISPR-driven editing at a second locus. GOI, gene of interest. (b) Typical selection process. Timing can vary according to initial modification rates at ATP1A1. (c) Schematic of the dual eSpCas9(1.1) and tandem U6-driven sgRNAs expression vector (see also Supplementary Figure 12). (d) Surveyor assays to determine on-target and off-target activity for the EMX1 sgRNA on samples reported in Table 1. (e) Same as (d) but for co-targeting AAVS1. (f) Surveyor assays on samples from K562 cells transiently co-transfected with WT SpCas9 and sgRNA expression vectors targeting AAVS1 (0.5μg and 1μg of each vector).
