Fig. 2: Investigation of on- and off-target mutations and determinants of off-target mutations.
From: Targeted base editing in the plastid genome of Arabidopsis thaliana
a, Mutated read frequencies in the plastid genomes of nine T1 lines (11 DAS) and one T2 line (49 DAS) targeted for 16S rRNA. SNPs where ≥10% of the reads were different from the reference genome (AP000423.1) in at least one plant are listed. Supplementary Table 2 shows all the on- and off-target mutations in both organelle genomes, where different reads frequencies were ≥1%. b–e, Positions and frequencies of on- and off-target mutations (shown as green dots) in T1 plants targeting 16S rRNA (b), rpoC1 (d) and psbA (e) and in null-segregant T2 plants mutated in 16S rRNA (c). Magenta lines represent the target windows. The right panels are magnified views of the regions surrounded by the dotted rectangles in left panels within 1,000 bp from the target nucleotides in each gene (G5 in 16S rRNA, G3 in rpoC1 and C10 in psbA). f, List of the off-target mutations categorized by types. Off-target mutations within 2 kb of the target windows and/or within 20 bp of sequences similar to those recognized by TALEs and in other regions are shown. In b–f, off-target mutations were defined as SNPs in which C/G-to-T/A substitutions (allele frequencies ≥0.01) were detected only in the T1 or T2 plants but not in three wild-type plants used as controls.
