RRC ID 53507
Author Wheway G, Schmidts M, Mans DA, Szymanska K, Nguyen TT, Racher H, Phelps IG, Toedt G, Kennedy J, Wunderlich KA, Sorusch N, Abdelhamed ZA, Natarajan S, Herridge W, van Reeuwijk J, Horn N, Boldt K, Parry DA, Letteboer SJF, Roosing S, Adams M, Bell SM, Bond J, Higgins J, Morrison EE, Tomlinson DC, Slaats GG, van Dam TJP, Huang L, Kessler K, Giessl A, Logan CV, Boyle EA, Shendure J, Anazi S, Aldahmesh M, Al Hazzaa S, Hegele RA, Ober C, Frosk P, Mhanni AA, Chodirker BN, Chudley AE, Lamont R, Bernier FP, Beaulieu CL, Gordon P, Pon RT, Donahue C, Barkovich AJ, Wolf L, Toomes C, Thiel CT, Boycott KM, McKibbin M, Inglehearn CF, UK10K Consortium, University of Washington Center for Mendelian Genomics, Stewart F, Omran H, Huynen MA, Sergouniotis PI, Alkuraya FS, Parboosingh JS, Innes AM, Willoughby CE, Giles RH, Webster AR, Ueffing M, Blacque O, Gleeson JG, Wolfrum U, Beales PL, Gibson T, Doherty D, Mitchison HM, Roepman R, Johnson CA.
Title An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes.
Journal Nat Cell Biol
Abstract 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.
Volume 17(8)
Pages 1074-1087
Published 2015-8-1
DOI 10.1038/ncb3201
PMID 26167768
PMC PMC4536769
MeSH 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 Cytoskeletal Proteins 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
IF 17.728
Times Cited 89