Figure 3
From: Highly specific targeted mutagenesis in plants using Staphylococcus aureus Cas9
Targeted mutagenesis efficiency in target gene with canonical or non-canonical PAM for SaCas9.
(a,b) Heteroduplex mobility assay of SaCas9 expression calli. CYP72A33 and VIP1 target sequences have canonical PAM (5′-NNGRRT-3′) (upper panel). CYP72A32 and VIP1-like target sequences have non-canonical PAM (5′-NNGRRC-3′ and 5′-NNGRRA-3′) (lower panel). (c) Mutations detected in OsCYP72A33 and OsCYP72A32 gene loci. The wild-type sequence is shown at the top with the target sequence underlined and the PAM in bold. DNA deletions and insertions are shown as dashes and lower case red letters. The net change in length and the number of clones are noted to the right of each sequence (+, insertion; −, deletion; ×, number of clones). The asterisk (*) indicates the difference from the canonical PAM for SaCas9.
