Fig. 1

Fig. 1

CRISPR/Cas9-induced mutations in the zebrafish mc2r gene. (A) Diagram of the mc2r single-exon gene. The coding region (CDS) is depicted as white boxes, whereas the 5′ UTR and 3′ UTR are shown as black boxes. The target site is represented by a black arrow. (B) Sequence of induced deletions in the mc2r locus. The first line represents the sequence of a WT zebrafish strain. The target sequence is highlighted in red. The black arrow indicates the protospacer-adjacent motif (PAM). The second and third lines show the different mutations that were induced. The second and third lines show the different mutations that were induced, scoring the deletion of each mutation on the right side. (C) Amino acid sequence of the complete protein of the zebrafish mc2r gene. The first line is the sequence of a WT fish, and the second and third lines represent the predicted sequences of both mutations. The gray background indicates the conserved sequence between the mutated and WT strains. Asterisks represent a codon stop. (D) Effects of CRISPR/Cas9-mc2r on mc2r mRNA. RT‒qPCR analysis of mc2r mRNA levels in wild-type (WT) (white bar) and mc2r knockout (mc2r^KO) (black bar) fish. The results were normalized to β-actin (actb) and are expressed as the means ± SDs (n = 5 per group). *P < 0,05.