Fig. 1: SpRY Cas9 in combination with different deaminases enables efficient disruption of targeted splice acceptors.
From: SPLICER: a highly efficient base editing toolbox that enables in vivo therapeutic exon skipping
A Schematic representation of the approach to disrupting splicing elements with near-PAMless ABEs (blue) or CBEs (green) by tiling eight sgRNAs per exon (Created in BioRender. Miskalis, A. (2024) https://BioRender.com/x92p416). Comparison of SpCas9 and SpRY Cas9 editing efficiency at the splice acceptors (SAs) of target exons using ABEs (B, C) or CBEs (D, E). Summary of editing efficiency at individual targets accomplished with SpCas9 or SpRY Cas9 ABEs (C) or CBEs (E). Comparison of DNA editing efficiency at multiple SAs using SpRY Cas9 fused with one of four different adenosine deaminases (F, G) or one of six different cytosine deaminases (H, I). Summary of editing efficiency at individual targets accomplished with different SpRY ABEs (G) or different SpRY CBEs (I). For (F−I), DNA editing rates were normalized to the protein expression of each deaminase from Fig. S1 by dividing each DNA editing rate by the normalized expression of each base editor. Values represent means and error bars indicate SD. For (B, D, F, and H), n = 2; n = 10 for (C); n = 9 for (E, G); n = 8 for (I). All replicates are biological replicates; ns, not significant; *p < 0.05, **p < 0.01; ***p < 0.001; two-tailed, unpaired t-test except for (G, I), which were analyzed via one-way ANOVA with Tukey’s post hoc. Source data are provided as a Source Data file.
