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; Rachel H  Giles

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, Rachel H Giles,

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

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)	1074-87
Number of pages	14
Journal	Nature Cell Biology
Volume	17
Issue number	8
DOIs	https://doi.org/10.1038/ncb3201
Publication status	Published - Aug 2015

Keywords

Abnormalities, Multiple
Animals
Caenorhabditis elegans
Cerebellar Diseases
Cerebellum
Cilia
Ciliary Motility Disorders
Databases, Genetic
Ellis-Van Creveld Syndrome
Eye Abnormalities
Genetic Markers
Genetic Predisposition to Disease
Genetic Testing
Genome-Wide Association Study
Genomics
HEK293 Cells
High-Throughput Nucleotide Sequencing
Humans
Kidney Diseases, Cystic
Membrane Proteins
Mice, Inbred C57BL
Mice, Knockout
Mutation
Phenotype
Photoreceptor Cells
Pregnancy Proteins
Proteins
RNA Interference
Reproducibility of Results
Retina
Suppressor Factors, Immunologic
Transfection
Zebrafish

Access to Document

10.1038/ncb3201

Cite this

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. (2015). An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes. Nature Cell Biology, 17(8), 1074-87. https://doi.org/10.1038/ncb3201

@article{e68a68c79339443db9c413f86ab312ee,

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.",

keywords = "Abnormalities, Multiple, Animals, Caenorhabditis elegans, Cerebellar Diseases, Cerebellum, Cilia, Ciliary Motility Disorders, Databases, Genetic, Ellis-Van Creveld Syndrome, Eye Abnormalities, Genetic Markers, Genetic Predisposition to Disease, Genetic Testing, Genome-Wide Association Study, Genomics, HEK293 Cells, High-Throughput Nucleotide Sequencing, Humans, Kidney Diseases, Cystic, Membrane Proteins, Mice, Inbred C57BL, Mice, Knockout, Mutation, Phenotype, Photoreceptor Cells, Pregnancy Proteins, Proteins, RNA Interference, Reproducibility of Results, Retina, Suppressor Factors, Immunologic, Transfection, Zebrafish",

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 Giles, {Rachel H}",

year = "2015",

month = aug,

doi = "10.1038/ncb3201",

language = "English",

volume = "17",

pages = "1074--87",

journal = "Nature Cell Biology",

issn = "1465-7392",

publisher = "Nature Research",

number = "8",

}

Wheway, G, Schmidts, M, Mans, DA, Szymanska, K, Nguyen, T-MT, 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, Giles, RH 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. 1074-87. 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 - Giles, Rachel H

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.

KW - Abnormalities, Multiple

KW - Animals

KW - Caenorhabditis elegans

KW - Cerebellar Diseases

KW - Cerebellum

KW - Cilia

KW - Ciliary Motility Disorders

KW - Databases, Genetic

KW - Ellis-Van Creveld Syndrome

KW - Eye Abnormalities

KW - Genetic Markers

KW - Genetic Predisposition to Disease

KW - Genetic Testing

KW - Genome-Wide Association Study

KW - Genomics

KW - HEK293 Cells

KW - High-Throughput Nucleotide Sequencing

KW - Humans

KW - Kidney Diseases, Cystic

KW - Membrane Proteins

KW - Mice, Inbred C57BL

KW - Mice, Knockout

KW - Mutation

KW - Phenotype

KW - Photoreceptor Cells

KW - Pregnancy Proteins

KW - Proteins

KW - RNA Interference

KW - Reproducibility of Results

KW - Retina

KW - Suppressor Factors, Immunologic

KW - Transfection

KW - Zebrafish

UR - http://www.scopus.com/inward/record.url?scp=84938681395&partnerID=8YFLogxK

U2 - 10.1038/ncb3201

DO - 10.1038/ncb3201

M3 - Article

C2 - 26167768

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

Keywords

Access to Document

Other files and links

Fingerprint

Cite this