Fig 1

(A) Schematic structure of zebrafish Cep290 illustrating the location of predicted protein domains. Domain structure is based on prior results [12, 44, 45]. The cep290^fh297 allele generates a premature stop codon at amino acid 1217. (B) Chromatograms of Sanger sequencing reactions of cDNAs from wild-type and homozygous cep290^fh297/fh297mutant confirming the C>T replacement. (C) Quantification of cep290 mRNA in 6 month old wild-type and mutant retinas by digital droplet PCR (ddPCR). (D) Lateral views of representative wild-type (top), heterozygous (middle), and homozygous (bottom) mutants at 5 dpf. At larval stage 30% of cep290^fh297/fh297 mutant animals show ventral tail curvature. (E) Lateral view of 7 month old wild-type, heterozygous and a representative cep290^fh297/fh297 mutants displaying distorted vertebral column. At adult stage 100% of the homozygous mutant animals show scoliosis of the vertebral column. (F) Representative micro-CT-generated images of lateral (top) and dorsal (bottom) views of adult wild-type and cep290^fh297/fh297 mutants. Images demonstrate that spinal curvature deviates within the dorsal/ventral plane as well as curves laterally (arrows). (G, top) Forward (F1) and reverse (R1) PCR primers were designed to bind sequences in exons 27 and 32 in order to flank exon 29 harboring the mutant cep290^fh297 allele. Exon skipping would result in a truncated PCR product of 593 bp, while retention of exon 29 would result in a full-length 1040 bp product. (G, bottom) Results of PCR reactions from cDNAs generated from wild-type and samples of two individual mutants resulted in full-length products of 1040 bp. 100 bp ladder shown in lane 1. (H) Optokinetic response (OKR) contrast response function of 5 dpf wild-type larvae (n = 11; open circles) and cep290^fh297/fh297 mutants (n = 26; closed circles). No significant differences were found. (I) OKR spatial frequency results for wild-type larvae (n = 13) and cep290^fh297/fh297mutants (n = 15). Error bars = s.e.m.