Fig. 2

Schematic diagram of OsGAD3 (Os03g0236200) gene structure and gRNA target sit for CRISPR/Cas9-mediated mutagenesis of OsGAD3. a Exons, introns, and UTRs are indicated by solid boxes, lines and blank boxes, respectively. Upstream and downstream arrows indicate translation initiation and termination sites, respectively. The numbers in brackets indicate the distance of an exon sequence to the initiation codon (ATG). A gray box indicates a calmodulin-binding domain (CaMBD). Scale corresponds to 100 bp. b Downstream region of OsGAD3 gene from nucleotide position 1366 to 1545. The amino acid sequence is shown below nucleotide sequence. An asterisk indicates a termination codon. Nucleotide sequence and its translated amino acid corresponded to a CaMBD is indicated in blue. Two target sequences (F1 target and R1 target) of the CRISPR/Cas9 are underlined. Boxes show the protospacer adjacent motif (PAM: 5′-NGG-3′) sequences. Black arrows show a putative cleavage sites of gRNA. Two termination codons are indicated in red. The former is an authentic termination codon and the latter one with a line is used after precious cleavage at the two cleavage sites and joining