Fig. 1

Targeted regions (a) and expression vector (b) of simultaneous site-directed mutagenesis of soybean genes Gly m Bd 28 K and Gly m Bd 30 K by using the CRISPR/Cas9 system. a Locations of the targeted sites of Gly m Bd 28 K and Gly m Bd 30 K genes. Boxes and pentagons, exons of the genes; bold lines, introns of the genes. Red and blue nucleotide sequences denote the gRNAs-targeted regions and the proto-spacer adjacent motif (PAM) regions, respectively. b Vector structure of the CRISPR/Cas9 expression module for soybean transformation. Bar cassette, Bar-marker gene unit; PcUbi pro, parsley (Petroselinum crispum) ubiquitin promoter; AtHSP ter, terminator of an Arabidopsis heat shock protein gene; AtU6 pro, Arabidopsis U6 promoter; GmU6–16 g pro, soybean U6–16 promoter