Figure 5. Testing the mutagenic activity of gRNA arrays in cell culture.

(A) Mismatches to GFP classified by base substitution in Kc cells transfected with human act5C-AID123*-nCas9 and 4 (top) of 24 (bottom) gRNAs, 2 weeks post-transfection. (B) Mismatches to GFP classified by base substitution in Kc cells transfected with sea lamprey act5C-AID^evoCDA1-nCas9 and 4 (top) of 24 (bottom) gRNAs, 2 weeks post-transfection. (C) Average mutation profiles over all gRNA used in the 4 (top) of 24 (bottom) gRNAs condition with different mutagenic enzymes. 24 gRNAs (bottom) mutate along the whole 200bp sequence and, when mutation rates are averaged over all gRNAs, the obtained mutation profiles exhibit less of a 5’ bias than a smaller number of non overlapping gRNAs (top panels).