Fig. 4.

Generation of Tg[pax7a-hs:GFP] and Tg[sox5-hs:GFP] fish. (a) Schematic of the knock-in strategies. The sgRNAs for pax7a or sox5 were designed to target a sequence upstream of the initiation methionine that corresponds to the 5′ untranslated region. (b–d) Tg[pax7a-hs:GFP]. (e–g) Tg[sox5-hs:GFP]. (b, b', c, c', e, e', f, f') Dorsal views of the head (b, b', e, e') and the trunk (c, c', f, f') at 2 dpf in the bright field (b, c, e, f) and in fluorescence (b', c', e', f'). (d, g) Dorsal (upper) and lateral (lower) views at 9 dpf. In Tg[pax7a-hs:GFP] embryos, GFP is expressed in the tectum, hindbrain, anterior neural tube (b'), and muscles (c'). These fluorescent signals are maintained in the hatchlings (d). In Tg[sox5-hs:GFP] embryos, GFP is expressed in a range of the central nervous system (CNS) from the forebrain to hindbrain (e'), and in neural tubes and the premigratory neural crest in a dotted manner (f'). At 9 dpf, additional fluorescent signals are observed in the pectoral fins, olfactory bulbs, and presumable xanthophore progenitors on the dorsal body surface (g; see also Additional file 3: Fig. S1C–D)