FIGURE 4.

Identifying transgene-free homozygous slpg mutants [(A) slpg-T₃#2, (B) slpg-T₃#3, and (C) slpg-T₃#5]. Gel image of PCR products obtained with primer sets for four T-DNA regions of sgRNA/Cas9 vector. SlActin (300 bp), part of the tomato Actin coding sequence, was used as a normalization control; Cas9 (875 bp), part of the Cas9 coding sequence; Pro35S + Cas9 (690 bp), region from the downstream of the 35S promoter to the upstream of the Cas9 coding sequence; SgRNA (639 bp), region from the AtU6 promoter to the downstream vector sequence spanning the sgRNA; Kanamycin (752 bp), part of the nptII coding sequence; M, DL5000 ladder DNA marker; WT, DNA of WT plant as a template; Lane 1-20, individual mutant lines; P, positive control (sgRNA/Cas9 vector plasmid as template); A, positive control (Agrobacterium containing sgRNA/Cas9 vector as template); N, negative control (water as template).