Fig. 1: Characterization of DddAtox variants derived from structure-based screening.
From: Base editing in human cells with monomeric DddA-TALE fusion deaminases
a Crystal structure of DddAtox (gray, PDB 6U08). The positively charged amino-acid residues and E1347 active site are shown in blue and orange, respectively. K1424 is not shown because 1423–1427 residues are not resolved in the currently available crystal structure. green; Zn2+ ion. b Graph showing the relative proportions of E. coli transformants obtained for each of the DddAtox variants containing alanine substitutions. The variant containing E1347A, a mutation affecting the active site, was used as a control. c, d Editing and indel frequencies induced by the indicated DddAtox AAAAA variant and E1347A variant fusions (c) and CBEs (d) at the TYRO3 site. e Heat map showing the frequencies of C-to-T substitutions at various positions in the TYRO3 site. The cytosine conversion rates and indel frequencies were measured by targeted deep sequencing. The protospacer is shown in blue and the PAM in orange. Cytosines upstream of the protospacer that underwent editing are shown in red. The numbers used to indicate the position of the DddA target window were obtained by counting backward from the proto-spacer, toward the 5′ upstream regions. Means ± s.e.m. (b–d) and heat map colors (e) were determined from three independent experiments. Source data are provided with this paper.
