Induction of large deletions using co-transformation of two genome editing vectors.

(A) Design of gRNAs to the MpNOP1 locus to dissect efficiencies of induction of large deletions. Grey boxes and lines indicate exons and introns. The dotted line indicates the downstream region of MpNOP1. gRNA positions are shown in red lines. Primer sets for PCR-based genotyping are also shown. (B) Representative images of electrophoresis of PCR-based genotyping. The gRNAs and primer sets used in the genotyping are shown. Expected sizes of the PCR products from the wild-type genome are colored in blue. Expected sizes of the PCR products from the genome in which inductions of large deletion occurred are colored in magenta. PCR products from non-specific amplifications are indicated by asterisks. (C) Summary table of the co-transformation experiment using pMpGE013 and pMpGE014. The ratio in the “Large deletion” row shows the number of T1 plants harboring the expected large deletion out of all the T1 plants inspected.