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

An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes
An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes
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.
1465-7392
1074-1087
Wheway, Gabrielle
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Schmidts, Miriam
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Mans, Dorus A.
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Szymanska, Katarzyna
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Nguyen, Thanh-Minh T.
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Racher, Hilary
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Phelps, Ian G.
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Wheway, Gabrielle
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Mans, Dorus A.
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Doherty, Dan
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Wheway, Gabrielle, Schmidts, Miriam, Mans, Dorus A., Szymanska, Katarzyna, Nguyen, Thanh-Minh T., Racher, Hilary, Phelps, Ian G., Toedt, Grischa, Kennedy, Julie, Wunderlich, Kirsten A., Sorusch, Nasrin, Abdelhamed, Zakia A., Natarajan, Subaashini, Herridge, Warren, Van Reeuwijk, Jeroen, Horn, Nicola, Boldt, Karsten, Parry, David A., Letteboer, Stef J. F., Roosing, Susanne, Adams, Matthew, Bell, Sandra M., Bond, Jacquelyn, Higgins, Julie, Morrison, Ewan E., Tomlinson, Darren C., Slaats, Gisela G., Van Dam, Teunis J. P., Huang, Lijia, Kessler, Kristin, Giessl, Andreas, Logan, Clare V., Boyle, Evan A., Shendure, Jay, Anazi, Shamsa, Aldahmesh, Mohammed, Al Hazzaa, Selwa, Hegele, Robert A., Ober, Carole, Frosk, Patrick, Mhanni, Aizeddin A., Chodirker, Bernard N., Chudley, Albert E., Lamont, Ryan, Bernier, Francois P., Beaulieu, Chandree L., Gordon, Paul, Pon, Richard T., Donahue, Clem, Barkovich, A. James, Wolf, Louis, Toomes, Carmel, Thiel, Christian T., Boycott, Kym M., Mckibbin, Martin, Inglehearn, Chris F., Stewart, Fiona, Omran, Heymut, Huynen, Martijn A., Sergouniotis, Panagiotis I., Alkuraya, Fowzan S., Parboosingh, Jillian S., Innes, A. Micheil, Willoughby, Colin E., Giles, Rachel H., Webster, Andrew R., Ueffing, Marius, Blacque, Oliver, Gleeson, Joseph G., Wolfrum, Uwe, Beales, Philip L., Gibson, Toby, Doherty, Dan, Mitchison, Hannah M., Roepman, Ronald and Johnson, Colin A. (2015) An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes. Nature Cell Biology, 17 (8), 1074-1087. (doi:10.1038/ncb3201).

Record type: Article

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.

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More information

Accepted/In Press date: 5 June 2015
e-pub ahead of print date: 13 July 2015
Published date: August 2015

Identifiers

Local EPrints ID: 423569
URI: http://eprints.soton.ac.uk/id/eprint/423569
ISSN: 1465-7392
PURE UUID: acbb42b2-a9aa-4475-a2bf-ac033f26f905
ORCID for Gabrielle Wheway: ORCID iD orcid.org/0000-0002-0494-0783

Catalogue record

Date deposited: 26 Sep 2018 16:30
Last modified: 28 Apr 2022 02:25

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Contributors

Author: Miriam Schmidts
Author: Dorus A. Mans
Author: Katarzyna Szymanska
Author: Thanh-Minh T. Nguyen
Author: Hilary Racher
Author: Ian G. Phelps
Author: Grischa Toedt
Author: Julie Kennedy
Author: Kirsten A. Wunderlich
Author: Nasrin Sorusch
Author: Zakia A. Abdelhamed
Author: Subaashini Natarajan
Author: Warren Herridge
Author: Jeroen Van Reeuwijk
Author: Nicola Horn
Author: Karsten Boldt
Author: David A. Parry
Author: Stef J. F. Letteboer
Author: Susanne Roosing
Author: Matthew Adams
Author: Sandra M. Bell
Author: Jacquelyn Bond
Author: Julie Higgins
Author: Ewan E. Morrison
Author: Darren C. Tomlinson
Author: Gisela G. Slaats
Author: Teunis J. P. Van Dam
Author: Lijia Huang
Author: Kristin Kessler
Author: Andreas Giessl
Author: Clare V. Logan
Author: Evan A. Boyle
Author: Jay Shendure
Author: Shamsa Anazi
Author: Mohammed Aldahmesh
Author: Selwa Al Hazzaa
Author: Robert A. Hegele
Author: Carole Ober
Author: Patrick Frosk
Author: Aizeddin A. Mhanni
Author: Bernard N. Chodirker
Author: Albert E. Chudley
Author: Ryan Lamont
Author: Francois P. Bernier
Author: Chandree L. Beaulieu
Author: Paul Gordon
Author: Richard T. Pon
Author: Clem Donahue
Author: A. James Barkovich
Author: Louis Wolf
Author: Carmel Toomes
Author: Christian T. Thiel
Author: Kym M. Boycott
Author: Martin Mckibbin
Author: Chris F. Inglehearn
Author: Fiona Stewart
Author: Heymut Omran
Author: Martijn A. Huynen
Author: Panagiotis I. Sergouniotis
Author: Fowzan S. Alkuraya
Author: Jillian S. Parboosingh
Author: A. Micheil Innes
Author: Colin E. Willoughby
Author: Rachel H. Giles
Author: Andrew R. Webster
Author: Marius Ueffing
Author: Oliver Blacque
Author: Joseph G. Gleeson
Author: Uwe Wolfrum
Author: Philip L. Beales
Author: Toby Gibson
Author: Dan Doherty
Author: Hannah M. Mitchison
Author: Ronald Roepman
Author: Colin A. Johnson

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