Fig. 4
From: Harnessing A3G for efficient and selective C-to-T conversion at C-rich sequences
Cas9-independent editing on DNA. The principle of the assay is illustrated at the left using oA3G-BE4max as an example, whereas the results are shown as bar graphs at the right. dSaCas9 recruited by a gRNA generates an R-loop at the specified genomic site (sites 5 and 6 in the current study; top left). A subset of the C’s within the R-loop is potentially susceptible to stochastic deamination by oA3G carried in oA3G-BE4max. On-target editing (at the EMX1-2) is induced to serve as a positive control for the off-target editing (bottom left). dSaCas9 and its gRNAs targeting site 5 or site 6 were coexpressed in HEK293T cells with the indicated editors and the gRNA targeting EMX1-2, and the editing analyzed 3 days later by Sanger sequencing. For on-target editing, only the data that controlled for site 5 editing is shown; the data for site 6 is highly similar and omitted for clarity. Values in the bar graphs are mean ± SEM (n = 3). Sanger chromatograms are shown in Additional file 4: Fig. S4