Fig. 5: Comparison of ME-ABEs with alternative engineered precision editors.

a, The subset of editors analyzed for precision editing are shown, highlighting derivation ancestry. Residues that differentiate each precision editor from their parent editor are listed—those that correspond to the wtTadA identity (reversions) are underlined, those that have been modified multiple times over the course of engineering (‘evolutionary hot spots’) are shown in bold, and all others are in plain text. b, HEK293T cells were transfected with plasmids encoding one of the listed ABEs and one of 12 gRNAs targeting an endogenous site or an NT control (Supplementary Fig. 18; indicated by asterisks). After a 72-h incubation, genomic DNA was collected, target sites were amplified and sequences were analyzed by NGS. Editing was performed in n = 3 biological replicates and normalized as described in Fig. 4. Normalized editing scores for all 96 A’s were then sorted by A position, and scores were further averaged across gRNAs to identify the editing window. The number of A’s considered for each position is given below the x-axis. The horizontal dotted line shows the 15% cutoff used to determine the editing window for each editor. c–g, Data from b are shown for editors ABE7.10 (c), ABE7.10-HRHSK (d), ABE8e (e), ABE8e + V106W (f) and ABE9 (g). Points show normalized editing efficiency for a given editor at a given site and are shown for all replicates. Box hinges illustrate quartiles, lines show medians and bars show minimum and maximum editing. h, Orthogonal R-loop assay was used to analyze gRNA-independent off-target activity of precision editors as described in Fig. 4d. The x–y plot indicates the average off-target activity for the most edited A position across three independent off-target sites, and average on-target activity at the most edited A position across two independent on-target sites, accounting for n = 3 biological replicates with error bars showing propagated standard deviation.

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