Cep290 disruption causes ciliopathy in zebrafish. (A) RT-PCR from representative single wild-type (WT) and cep290ex25 morphant embryos. Cep290 morpholino oligonucleotide targeted to the exon 25 splice donor caused mispliced RNAs and intron retention. (B) Chromatogram of a CRISPR/Cas9-generated single representative genetic cep290 mutant showing the 10-bp deletion in exon 16 that results in a premature stop codon. (C) Morphology of an uninjected embryo compared to cep290ex25 and cep290ATG morphants at 2 dpf. (D) Morphology of wild-type and MZcep290fb208/fb208 mutants (hereafter called MZcep290−/− mutants) at 3 dpf. (E) Western blot showing loss of Cep290CT immunoreactivity in cep290ATG and cep290ex25 morphants (pool of 40 48-hpf embryos). The position of the 250-kDa molecular weight marker is indicated in red. A tubulin antibody was used as a loading control (bottom). (F) Western blot showing loss of Cep290CT and Cep290NT immunoreactivity in MZcep290−/− mutants. A tubulin antibody was used as a loading control. Experiments were replicated a minimum of three times.
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