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

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

doi:10.1038/ncb3201

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

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

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

186 Citations (Scopus)

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.

Original language	English
Pages (from-to)	1074-1087
Number of pages	14
Journal	Nature Cell Biology
Volume	17
Issue number	8
Early online date	13 Jul 2015
DOIs	https://doi.org/10.1038/ncb3201
Publication status	Published - Aug 2015

ASJC Scopus subject areas

Cell Biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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Wheway, G., Schmidts, M., Mans, D. A., Szymanska, K., Nguyen, T. M. T., 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. F., ... Johnson, C. 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. https://doi.org/10.1038/ncb3201

@article{93c278a036f34c05a5d6d2e3a8530251,

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

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.",

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

year = "2015",

month = aug,

doi = "10.1038/ncb3201",

language = "English",

volume = "17",

pages = "1074--1087",

journal = "Nature Cell Biology",

issn = "1465-7392",

publisher = "Nature Research",

number = "8",

}

Wheway, G, Schmidts, M, Mans, DA, Szymanska, K, Nguyen, TMT, 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, 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 2015, 'An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes', Nature Cell Biology, vol. 17, no. 8, pp. 1074-1087. https://doi.org/10.1038/ncb3201

TY - JOUR

T1 - An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes

AU - Wheway, Gabrielle

AU - Schmidts, Miriam

AU - Mans, Dorus A.

AU - Szymanska, Katarzyna

AU - Nguyen, Thanh Minh T

AU - Racher, Hilary

AU - Phelps, Ian G.

AU - Toedt, Grischa

AU - Kennedy, Julie

AU - Wunderlich, Kirsten A.

AU - Sorusch, Nasrin

AU - Abdelhamed, Zakia A.

AU - Natarajan, Subaashini

AU - Herridge, Warren

AU - Van Reeuwijk, Jeroen

AU - Horn, Nicola

AU - Boldt, Karsten

AU - Parry, David A.

AU - Letteboer, Stef J F

AU - Roosing, Susanne

AU - Adams, Matthew

AU - Bell, Sandra M.

AU - Bond, Jacquelyn

AU - Higgins, Julie

AU - Morrison, Ewan E.

AU - Tomlinson, Darren C.

AU - Slaats, Gisela G.

AU - Van Dam, Teunis J P

AU - Huang, Lijia

AU - Kessler, Kristin

AU - Giessl, Andreas

AU - Logan, Clare V.

AU - Boyle, Evan A.

AU - Shendure, Jay

AU - Anazi, Shamsa

AU - Aldahmesh, Mohammed

AU - Al Hazzaa, Selwa

AU - Hegele, Robert A.

AU - Ober, Carole

AU - Frosk, Patrick

AU - Mhanni, Aizeddin A.

AU - Chodirker, Bernard N.

AU - Chudley, Albert E.

AU - Lamont, Ryan

AU - Bernier, Francois P.

AU - Beaulieu, Chandree L.

AU - Gordon, Paul

AU - Pon, Richard T.

AU - Donahue, Clem

AU - Barkovich, A. James

AU - Wolf, Louis

AU - Toomes, Carmel

AU - Thiel, Christian T.

AU - Boycott, Kym M.

AU - McKibbin, Martin

AU - Inglehearn, Chris F.

AU - Stewart, Fiona

AU - Omran, Heymut

AU - Huynen, Martijn A.

AU - Sergouniotis, Panagiotis I.

AU - Alkuraya, Fowzan S.

AU - Parboosingh, Jillian S.

AU - Innes, A. Micheil

AU - Willoughby, Colin E.

AU - Giles, Rachel H.

AU - Webster, Andrew R.

AU - Ueffing, Marius

AU - Blacque, Oliver

AU - Gleeson, Joseph G.

AU - Wolfrum, Uwe

AU - Beales, Philip L.

AU - Gibson, Toby

AU - Doherty, Dan

AU - Mitchison, Hannah M.

AU - Roepman, Ronald

AU - Johnson, Colin A.

PY - 2015/8

Y1 - 2015/8

N2 - 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.

AB - 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.

U2 - 10.1038/ncb3201

DO - 10.1038/ncb3201

M3 - Article

AN - SCOPUS:84938681395

SN - 1465-7392

VL - 17

SP - 1074

EP - 1087

JO - Nature Cell Biology

JF - Nature Cell Biology

IS - 8

ER -

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

Abstract

ASJC Scopus subject areas

UN SDGs

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