Fig. 3
From: Genome-wide profiling of adenine base editor specificity by EndoV-seq
Using multiplex EndoV-seq to profile off-target effects of ABE. a Human genomic DNA (10 μg) was treated with a mixture of 300 nM ABE7.10 and six gRNAs (200 nM each) (HEK293-2, EMX1, FANCF, HBB-28 (T > C), RNF2 and HBG) for 8 h and then with EndoV (8 U) for 3 h. The treated gnomic DNA was subsequently sequenced with ~30–40-fold coverage and genome-wide cleavage scores calculated. Sites with cleavage score of >2.5 are plotted here. Orange, genomic DNA treated with ABE7.10 and multiplex gRNAs. Gray, untreated genomic DNA. b Genome-wide cleavage scores (cutoff > 2.5) of untreated (gray), monoplex EndoV-seq (blue), and multiplex EndoV-seq (orange) for the indicated gRNAs are plotted. Red arrows, on-target sites. c Sequence logos (by WebLogo) of multiplex EndoV-seq captured off-target (DNA cleavage scores of >2.5) and on-target sites. d A comparison of monoplex vs. multiplex EndoV-seq captured off-target sites (DNA cleavage scores of >2.5, on-target sites not shown)
