Fig. 6: gRNA and target properties affect mutagenesis efficiency.

a Since Target-AID can generate both C to G and C to T mutations, many codons can be targeted to create premature stop codons. The TGG (W) codon is the only one targeted on the non-coding strand as ACC. b GNE ratio for SGGs targeting different codons in essential genes, split by co-editing risk categories, where 1 and 2 represent low or very low co-editing risk while 3 or 4 represent moderate to high co-editing risk. c Cumulative z-score density of SGGs grouped by the mutational outcome generating the stop codon. A higher rate of GNE is observed for gRNAs for which a C-to-G mutation at the highest editing activity position generates a stop codon mutation. The significance threshold is shown as a black dotted line. d Cumulative z-score density of gRNAs that do not generate stop codons targeting either the coding or non-coding strand. e SGG and non-SGG GNE enrichment compared to the expected GNE ratio for different melting temperature ranges. f GC, C, and G content of NSGs and GNEs. Distribution medians are shown as black dotted lines and means are shown as red lines. P-values were calculated using Welch’s t-tests. Boxplots represent the upper and lower quartiles of the data, and whiskers extend to 1.5 times the interquartile range (Q3–Q1) at most. Outliers are shown in gray.