Fig. 2: Characterization of the DddAtox GSVG variant derived from random mutagenesis.
From: Base editing in human cells with monomeric DddA-TALE fusion deaminases
a Schematic diagram of the screen for non-toxic DddAtox variants generated by error-prone PCR. The E1347 active site and replaced residues are shown in violet and red, respectively. b, c Editing and indel frequencies at the TYRO3 site induced by the GSVG variant fused to the N (b) and C (c) termini of Cas9, D10A nCas9, H840A nCas9, and D10A, H840A dCas9 in HEK293T cells. d, Heat map showing the frequencies of C-to-T substitutions at various positions in the TYRO3 site. The protospacer is shown in blue and the PAM in orange. Cytosines that underwent editing are shown in red. the N terminus and C terminus of Cas9, nCas9(D10A), nCas9(H840A), and dCas9, respectively. The numbers used to indicate the position of the GSVG target window were obtained by counting backward and forward from the proto-spacer, toward the 5′ upstream and toward the 3′ downstream regions, respectively. The base editing and indel frequencies were measured by targeted deep sequencing. Means ± s.e.m. (b, c) and heat map colors (d) were determined from three independent experiments. Source data are provided with this paper.
