Supplementary Figure 6: Properties of base editors derived from enAsCas12a and Cas12a orthologs.

(a) Schematic of dCas12a base-editor (BE) constructs with varying NLS and linker compositions. NLS(sv), SV40 nuclear localization signal; NLS(nuc), nucleoplasmin nuclear localization signal; gs, glycine-serine peptide linker; rAPO1, rat APOBEC1; UGI, uracil glycosylase inhibitor. (b) Fold-change in cytosine to thymine (C-to-T) editing compared to the untreated control across all Cs in the 20 nt spacers of 8 target sites. (c) Influence of identity of the preceding (5’) base on C-to-T editing efficiency across eight target sites (see Fig. 3h) is plotted for all Cs in the window encompassing the -14 to +30 region of each target site (an additional 10 nt upstream of the 4 nt PAM and 10 nt downstream of the 20 nt spacer sequence; see Supplementary Table 3). (d) Analysis of edit purity at six selected cytosines across five target sites. The fraction of each non-C identity is plotted over the sum of all non-C occurrences at that position for each BE construct. Bkgd, distribution of nucleotide substitutions or deletions in control samples. Mean and s.e.m. shown for n = 3. (e) Percent insertion or deletion mutation (indel) across sites targeted with dCas12a-BEs. Indels were calculated for each BE/crRNA pair by determining the percentage of alleles encoding an indel within the -14 to +30 window, not counting alleles with substitutions only. Mean and s.e.m. shown for n = 3.