Figure 3

Generation of c3orf70-KO zebrafish. (A) Heteroduplex mobility assay of PCR products after amplification of the crRNA-targeted genomic region of c3orf70a and c3orf70b. Genomic DNA was extracted from c3orf70 wild-type (WT) and c3orf70-KO zebrafish and subjected to PCR to amplify short fragments of the c3orf70a and c3orf70b genes, including their crRNA target sites. The PCR products were electrophoresed in a 10% acrylamide gel. The positions of the expected homoduplexes for WT and KO zebrafish are indicated by black and red arrowheads, respectively. (B) Nucleotide sequence alignment of c3orf70a and c3orf70b genes from WT and KO zebrafish. The recognition sites of crRNA targeting c3orf70a and c3orf70b, including the PAM sequences, are shown in magenta and red boxes, respectively. (C) Alignment of amino acid sequences inferred from cDNA of c3orf70a and c3orf70b from WT and KO zebrafish.