Extended Data Fig. 3. Mutation table and representative activity of ePACE4 evolved Nme2Cas9 variants.

(a) Genotypes of individually sequenced plaques following ePACE4, with positions varying from wild-type displayed. Clones evolved on different PAMs are delineated by a bold line. Mutations that had previously appeared in ePACE1 and ePACE2 are shown in light pink and magenta, respectively, while novel mutations are shown in blue. (b) Heat map showing ABE-PPA activity of representative clones from ePACE4 on the 16 combinations of PAM positions 5 and 6 (N₄NN) Values are raw % A•T-to-G•C conversion observed for one replicate of each editor and are listed in each cell for the N₄CN PAMs, with values above 70% A•T-to-G•C conversion colored white. (c) ABE-PPA activity in (b) pooled and segregated by mutation position. Each column depicts the impact of a given position, when mutated, on ABE-PPA activity at each of the four PAM groups (N5A, N5C, N5G, N5T) (see Supplementary Note 4). Values are normalized against the highest activity within each set of PAMs. Only positions that were observed to be mutated more than once in (a) were included in this analysis.