The N-terminal region of centrosomal protein 290 (CEP290) restores vision in a zebrafish model of human blindness

Hum Mol Genet. 2011 Apr 15;20(8):1467-77. doi: 10.1093/hmg/ddr025. Epub 2011 Jan 21.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Animals, Genetically Modified
  • Antigens, Neoplasm / biosynthesis*
  • Antigens, Neoplasm / genetics
  • Blindness / genetics*
  • Cell Cycle Proteins
  • Cytoskeletal Proteins
  • Disease Models, Animal
  • Eye / embryology
  • Eye / metabolism
  • Eye Abnormalities / genetics
  • Gene Expression Regulation, Developmental
  • HEK293 Cells
  • Humans
  • Microtubule-Associated Proteins / genetics*
  • Microtubule-Associated Proteins / metabolism
  • Molecular Sequence Data
  • Mutation, Missense
  • Neoplasm Proteins / biosynthesis*
  • Neoplasm Proteins / genetics
  • Optic Atrophy, Hereditary, Leber / genetics
  • Phenotype
  • Protein Structure, Tertiary
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / genetics
  • Reflex, Startle
  • Tail / embryology
  • Transcription, Genetic
  • Vision, Ocular / genetics*
  • Zebrafish / embryology*
  • Zebrafish / genetics
  • Zebrafish / metabolism
  • Zebrafish Proteins / genetics*
  • Zebrafish Proteins / metabolism

Substances

  • Antigens, Neoplasm
  • CEP290 protein, zebrafish
  • Cell Cycle Proteins
  • Cep290 protein, human
  • Cytoskeletal Proteins
  • Microtubule-Associated Proteins
  • Neoplasm Proteins
  • Recombinant Proteins
  • Zebrafish Proteins