Split-SaCas9 transiently expressed separately via ToMV inoculation and Agroinfiltration induced targeted mutagenesis in N. benthamiana. (A) Schematic representation of the ToMV vector expressing full-length SaCas9 or split-SaCas9. A circle represents the cap structure, and rectangular boxes represent open reading frames. The 130K and 180K are involved in viral RNA replication. The 30K is involved in cell to cell movement. (B) Experimental scheme of ToMV infection and Agroinfiltration for genome editing. (C) Genomic DNA was extracted from leaves at 11 d after ToMV inoculation (7 d after Agroinfiltration). CAPS analysis of the sgPDS1 locus was performed as in Fig. 2A. +: BstNI-digested PCR products.