title: Species-dependent splice recognition of a cryptic exon resulting from a recurrent intronic CEP290 mutation that causes congenital blindness.
abstract: A mutation in intron 26 of CEP290 (c.2991+1655A>G) is the most common genetic cause of Leber congenital amaurosis (LCA), a severe type of inherited retinal degeneration. This mutation creates a cryptic splice donor site, resulting in the insertion of an aberrant exon (exon X) into ~50% of all CEP290 transcripts. A humanized mouse model with this mutation did not recapitulate the aberrant CEP290 splicing observed in LCA patients, suggesting differential recognition of cryptic splice sites between species. To further assess this phenomenon, we generated two CEP290 minigene constructs, with and without the intronic mutation, and transfected these in cell lines of various species. RT-PCR analysis revealed that exon X is well recognized by the splicing machinery in human and non-human primate cell lines. Intriguingly, this recognition decreases in cell lines derived from species such as dog and rodents, and it is completely absent in Drosophila. In addition, other cryptic splicing events corresponding to sequences in intron 26 of CEP290 were observed to varying degrees in the different cell lines. Together, these results highlight the complexity of splice site recognition among different species, and show that care is warranted when generating animal models to mimic splice site mutations in vivo.
authors.affiliation: Department of Human Genetics, Radboud University Medical Center, Geert Grooteplein Zuid 10, 6525 GA Nijmegen, The Netherlands. alex.garantoiglesias@radboudumc.nl.|Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands. alex.garantoiglesias@radboudumc.nl.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Garanto
authors.affiliation: Department of Human Genetics, Radboud University Medical Center, Geert Grooteplein Zuid 10, 6525 GA Nijmegen, The Netherlands. lonneke.duijkers@radboudumc.nl.
authors.forename: Lonneke
authors.initials: L
authors.lastname: Duijkers
authors.affiliation: Department of Human Genetics, Radboud University Medical Center, Geert Grooteplein Zuid 10, 6525 GA Nijmegen, The Netherlands. rob.collin@radboudumc.nl.|Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands. rob.collin@radboudumc.nl.
authors.forename: Rob W J
authors.initials: RW
authors.lastname: Collin
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm; D001483:Base Sequence; D018797:Cell Cycle Proteins; D002460:Cell Line; D003598:Cytoskeletal Proteins; D004285:Dogs; D004330:Drosophila / Q000235:genetics; D005091:Exons; D057809:HEK293 Cells; D006801:Humans; D007438:Introns; D057130:Leber Congenital Amaurosis / Q000235:genetics* / Q000473:pathology; D051379:Mice; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D012326:RNA Splicing; D013045:Species Specificity
chemical_list: D000951:Antigens, Neoplasm; D018797:Cell Cycle Proteins; C513600:Cep290 protein, mouse; D003598:Cytoskeletal Proteins; D009687:Nuclear Proteins