MZcep290−/− mutants present milder phenotypes that recover after 4-5 dpf. (A) Clutch of cep290 maternal zygotic cep290 mutants. Body shape curvature seen at 3 dpf was lost at 7 dpf (n=100). (B) Maternal zygotic cep290 mutants reach adulthood and can present scoliosis (15-40% of adult fish; n=31). (C) RT-PCR from MZcep290−/− and wild-type embryos. No exon skipping around exon 16 was detected in mutant embryos. (D) RT-qPCR from adult eye and kidney samples (n=3). Specific wild-type (WT) and mutant primers for the targeted exon 16 were used to quantify wild-type and mutated cep290 mRNA. As expected, wild-type primers did not generate an RT-PCR product with mutant mRNA samples relative to wild-type mRNA, whereas mutant primers generated significant product with mutant mRNA samples relative to wild-type mRNA samples. Primers for ex3-ex5, ex19-ex20 and ex50-52 were used to quantify nonsense-mediated decay. No changes in cep290 mRNA levels were detected in MZcep290−/− mutant samples compared with controls. n=3 biological replicates. Data are mean±s.d. Experiments were replicated at a minimum three times. *P<0.05; ****P<0.0001; ns, not significant (unpaired t-test).
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