The N-terminal region of centrosomal protein 290 (CEP290) restores vision in a zebrafish model of human blindness.
The gene coding for centrosomal protein 290 (CEP290), a large multidomain protein, is the most frequently mutated gene underlying the non-syndromic blinding disorder Leber's congenital amaurosis (LCA). CEP290 has also been implicated in several cilia-related syndromic disorders including Meckel-Gruber syndrome, Joubert syndrome, Senor-Loken syndrome and Bardet-Biedl syndrome (BBS). In this study, we characterize the developmental and functional roles of cep290 in zebrafish. An antisense oligonucleotide [Morpholino (MO)], designed to generate an altered cep290 splice product that models the most common LCA mutation, was used for gene knockdown. We show that cep290 MO-injected embryos have reduced Kupffer's vesicle size and delays in melanosome transport, two phenotypes that are observed upon knockdown of bbs genes in zebrafish. Consistent with a role in cilia function, the cep290 MO-injected embryos exhibited a curved body axis. Patients with LCA caused by mutations in CEP290 have reduced visual perception, although they present with a fully laminated retina. Similarly, the histological examination of retinas from cep290 MO-injected zebrafish revealed no gross lamination defects, yet the embryos had a statistically significant reduction in visual function. Finally, we demonstrate that the vision impairment caused by the disruption of cep290 can be rescued by expressing only the N-terminal region of the human CEP290 protein. These data reveal that a specific region of the CEP290 protein is sufficient to restore visual function and this region may be a viable gene therapy target for LCA patients with mutations in CEP290.
Department of Biology, University of Iowa, Iowa City, IA 52242, USA.
Lisa M
LM
Baye
Xiaobai
X
Patrinostro
Svetha
S
Swaminathan
John S
JS
Beck
Yan
Y
Zhang
Edwin M
EM
Stone
Val C
VC
Sheffield
Diane C
DC
Slusarski
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D000951:Antigens, Neoplasm / Q000096:biosynthesis* / Q000235:genetics; D001766:Blindness / Q000235:genetics*; D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D004195:Disease Models, Animal; D005123:Eye / Q000196:embryology / Q000378:metabolism; D005124:Eye Abnormalities / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D057809:HEK293 Cells; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics; D029242:Optic Atrophy, Hereditary, Leber / Q000235:genetics; D010641:Phenotype; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D013216:Reflex, Startle; D013623:Tail / Q000196:embryology; D014158:Transcription, Genetic; D014785:Vision, Ocular / Q000235:genetics*; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics / Q000378:metabolism; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism
D000951:Antigens, Neoplasm; C559615:CEP290 protein, zebrafish; D018797:Cell Cycle Proteins; C493903:Cep290 protein, human; D003598:Cytoskeletal Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D011994:Recombinant Proteins; D029961:Zebrafish Proteins
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