Generation of ict1 and mtrfr knockout zebrafish lines using the CRISPR/Cas9 system. (A) Upper panel: Schematic diagram of the configuration of ict1 from zebrafish according to the Ensemble database [56]. Exons are represented as boxes, with gray regions indicating the protein‐coding regions. The downward arrow indicates the mutation position. Only the DNA sequences of genes for the wild‐type (wt) and ict1−/− before and after the mutation position are shown, and the corresponding amino acid sequence for wt is shown above the DNA sequence. The guide RNA sequence in wt is underlined. The mutated sequence in ict1−/− is presented in bold, and the boxed sequence indicates a premature stop codon. Bottom panel: A schematic domain representation of the Ict1 protein. The structured catalytic domain (the GGQ domain) is represented as a box based on the mammal ICT1 structures. The gray bar indicates the region corresponding to a putative mitochondrial targeting sequence or presequence, which was predicted using the TargetP‐2.0 web server [57]. The upward arrow indicates the first amino acid that undergoes alteration due to the mutation. The amino acid sequences for the wt and ict1−/−are shown. Bold letters indicate mutated residues in ict1−/−, and an asterisk indicates termination. (B) The figure regarding mtrfr is shown. The notation is the same as (A).
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