An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes

Wheway, G., Schmidts, M., Mans, D.A., Szymanska, K., Nguyen, T.M., Racher, H., Phelps, I.G., Toedt, G., Kennedy, J., Wunderlich, K.A., Sorusch, N., Abdelhamed, Z.A., Natarajan, S., Herridge, W., van Reeuwijk, J., Horn, N., Boldt, K., Parry, D.A., Letteboer, S.J., Roosing, S., Adams, M., Bell, S.M., Bond, J., Higgins, J., Morrison, E.E., Tomlinson, D.C., Slaats, G.G., van Dam, T.J., Huang, L., Kessler, K., Giessl, A., Logan, C.V., Boyle, E.A., Shendure, J., Anazi, S., Aldahmesh, M., Al Hazzaa, S., Hegele, R.A., Ober, C., Frosk, P., Mhanni, A.A., Chodirker, B.N., Chudley, A.E., Lamont, R., Bernier, F.P., Beaulieu, C.L., Gordon, P., Pon, R.T., Donahue, C., Barkovich, A.J., Wolf, L., Toomes, C., Thiel, C.T., Boycott, K.M., McKibbin, M., Inglehearn, C.F., Stewart, F., Omran, H., Huynen, M.A., Sergouniotis, P.I., Alkuraya, F.S., Parboosingh, J.S., Innes, A.M., Willoughby, C.E., Giles, R.H., Webster, A.R., Ueffing, M., Blacque, O., Gleeson, J.G., Wolfrum, U., Beales, P.L., Gibson, T., Doherty, D., Mitchison, H.M., Roepman, R., Johnson, C.A.
Nature cell biology   17: 1074-87 (Journal)
Registered Authors
Lamont, Ryan, Phelps, Ian
Cilia, Ciliogenesis, RNAi
MeSH Terms
  • Abnormalities, Multiple
  • Animals
  • Caenorhabditis elegans/genetics
  • Caenorhabditis elegans/metabolism
  • Caenorhabditis elegans/ultrastructure
  • Cerebellar Diseases/genetics
  • Cerebellum/abnormalities
  • Cilia/genetics*
  • Cilia/metabolism
  • Cilia/pathology
  • Ciliary Motility Disorders/genetics*
  • Ciliary Motility Disorders/metabolism
  • Ciliary Motility Disorders/pathology
  • Databases, Genetic
  • Ellis-Van Creveld Syndrome/genetics
  • Eye Abnormalities/genetics
  • Genetic Markers*
  • Genetic Predisposition to Disease
  • Genetic Testing/methods*
  • Genome-Wide Association Study
  • Genomics/methods*
  • HEK293 Cells
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Kidney Diseases, Cystic/genetics
  • Membrane Proteins/deficiency
  • Membrane Proteins/genetics
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mutation
  • Phenotype
  • Photoreceptor Cells*/metabolism
  • Photoreceptor Cells*/ultrastructure
  • Pregnancy Proteins/genetics
  • Pregnancy Proteins/metabolism
  • Proteins/genetics
  • Proteins/metabolism
  • RNA Interference*
  • Reproducibility of Results
  • Retina/abnormalities
  • Suppressor Factors, Immunologic/genetics
  • Suppressor Factors, Immunologic/metabolism
  • Transfection
  • Zebrafish/genetics
  • Zebrafish/metabolism
26167768 Full text @ Nat. Cell Biol.
Defects in primary cilium biogenesis underlie the ciliopathies, a growing group of genetic disorders. We describe a whole-genome siRNA-based reverse genetics screen for defects in biogenesis and/or maintenance of the primary cilium, obtaining a global resource. We identify 112 candidate ciliogenesis and ciliopathy genes, including 44 components of the ubiquitin-proteasome system, 12 G-protein-coupled receptors, and 3 pre-mRNA processing factors (PRPF6, PRPF8 and PRPF31) mutated in autosomal dominant retinitis pigmentosa. The PRPFs localize to the connecting cilium, and PRPF8- and PRPF31-mutated cells have ciliary defects. Combining the screen with exome sequencing data identified recessive mutations in PIBF1, also known as CEP90, and C21orf2, also known as LRRC76, as causes of the ciliopathies Joubert and Jeune syndromes. Biochemical approaches place C21orf2 within key ciliopathy-associated protein modules, offering an explanation for the skeletal and retinal involvement observed in individuals with C21orf2 variants. Our global, unbiased approaches provide insights into ciliogenesis complexity and identify roles for unanticipated pathways in human genetic disease.
Genes / Markers
Show all Figures
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes