Cas9/sgRNA-induced mutations at the <i>Bm-ok</i> locus in G₀<i>Bombyx mori</i>.

<p>(A) Schematic representation of <i>Bm-ok</i>. exons are shown as boxes and arrows represent the primers used to amplify the target regions. Target site locations (<i>Bm-ok</i>-tar1 and <i>Bm-ok</i>-tar2) are underlined and PAM sequences are shown in red. (B) Representative chromatograms of PCR-product sequencing in G₀ silkworms in which indel mutations are present. (C) Sequences of indel mutations at the targeted <i>Bm-ok</i> locus in G₀ silkworms. The target sites are highlighted in green and PAM sequences are shown in red. Deletions are indicated by hyphens and insertions are shown in lowercase red letters. The indel mutation type is noted to the right (+, insertion; -, deletion). (D) Images indicating characteristic <i>Bm-ok</i> mosaic mutation phenotypes: (a) Fifth instar G₀ wild-type (P50) (left) and mosaic mutant (right) individuals; (b–d) Magnified images of wild-type and mutant individuals. The scale bars represent 1 mm.