|
Fig. 4
Biallelic disruption of golden (gol) by Cas9 generates mosaic pigmentation phenotypes. gol gRNA (50 pg) and nls-zCas9-nls RNA (150 pg) were injected into wild-type embryos. (A–F) Lateral views of wild-type (A and E) and gol-targeted embryos (B–D and F) at 2 dpf. Whereas the wild-type embryo showed darkly pigmented retinal pigment epithelium (RPE), gol-targeted embryos had patches of unpigmented cells in RPE (B–D). The skin melanophores in gol-targeted embryos were also hypopigmented (F). (G) T7EI assays showed high mutagenesis rates at the gol target (80–98%) in six randomly selected gol-targeted embryos. (H–K) Lateral views of wild-type (H and J) and gol-targeted (I and K) juvenile fish at 54 dpf. gol-targeted fish had patches of pale melanophores (dashed boxes) among the dark skin stripes. (Scale bars: in D, 300 μm; in F, 500 μm; in K, 2 mm.)