title: Accelerating Gene Discovery by Phenotyping Whole-Genome Sequenced Multi-mutation Strains and Using the Sequence Kernel Association Test (SKAT).
abstract: Forward genetic screens represent powerful, unbiased approaches to uncover novel components in any biological process. Such screens suffer from a major bottleneck, however, namely the cloning of corresponding genes causing the phenotypic variation. Reverse genetic screens have been employed as a way to circumvent this issue, but can often be limited in scope. Here we demonstrate an innovative approach to gene discovery. Using C. elegans as a model system, we used a whole-genome sequenced multi-mutation library, from the Million Mutation Project, together with the Sequence Kernel Association Test (SKAT), to rapidly screen for and identify genes associated with a phenotype of interest, namely defects in dye-filling of ciliated sensory neurons. Such anomalies in dye-filling are often associated with the disruption of cilia, organelles which in humans are implicated in sensory physiology (including vision, smell and hearing), development and disease. Beyond identifying several well characterised dye-filling genes, our approach uncovered three genes not previously linked to ciliated sensory neuron development or function. From these putative novel dye-filling genes, we confirmed the involvement of BGNT-1.1 in ciliated sensory neuron function and morphogenesis. BGNT-1.1 functions at the trans-Golgi network of sheath cells (glia) to influence dye-filling and cilium length, in a cell non-autonomous manner. Notably, BGNT-1.1 is the orthologue of human B3GNT1/B4GAT1, a glycosyltransferase associated with Walker-Warburg syndrome (WWS). WWS is a multigenic disorder characterised by muscular dystrophy as well as brain and eye anomalies. Together, our work unveils an effective and innovative approach to gene discovery, and provides the first evidence that B3GNT1-associated Walker-Warburg syndrome may be considered a ciliopathy.
authors: Department of Molecular Biology and Biochemistry and Centre for Cell Biology, Development, and Disease, Simon Fraser University, Burnaby, British Columbia, Canada.
authors: Tiffany A
authors: 0000-0002-2667-376X
authors: TA
authors: Timbers
authors: Department of Zoology, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Stephanie J
authors: SJ
authors: Garland
authors: Department of Molecular Biology and Biochemistry and Centre for Cell Biology, Development, and Disease, Simon Fraser University, Burnaby, British Columbia, Canada.
authors: Swetha
authors: S
authors: Mohan
authors: Department of Zoology, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Stephane
authors: S
authors: Flibotte
authors: Department of Zoology, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Mark
authors: 0000-0001-9113-8862
authors: M
authors: Edgley
authors: Department of Molecular Biology and Biochemistry and Centre for Cell Biology, Development, and Disease, Simon Fraser University, Burnaby, British Columbia, Canada.
authors: Quintin
authors: Q
authors: Muncaster
authors: Department of Zoology, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Vinci
authors: 0000-0002-3587-6200
authors: V
authors: Au
authors: Department of Zoology, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Erica
authors: 0000-0002-4361-151X
authors: E
authors: Li-Leger
authors: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Federico I
authors: 0000-0003-0680-4731
authors: FI
authors: Rosell
authors: Department of Molecular Biology and Biochemistry and Centre for Cell Biology, Development, and Disease, Simon Fraser University, Burnaby, British Columbia, Canada.
authors: Jerry
authors: J
authors: Cai
authors: Department of Cell Biology, Erasmus MC, Rotterdam, The Netherlands.
authors: Suzanne
authors: S
authors: Rademakers
authors: Department of Cell Biology, Erasmus MC, Rotterdam, The Netherlands.
authors: Gert
authors: 0000-0002-7524-171X
authors: G
authors: Jansen
authors: Department of Zoology, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Donald G
authors: DG
authors: Moerman
authors: Department of Molecular Biology and Biochemistry and Centre for Cell Biology, Development, and Disease, Simon Fraser University, Burnaby, British Columbia, Canada.
authors: Michel R
authors: MR
authors: Leroux
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism / Q000473:pathology; D017173:Caenorhabditis elegans / Q000235:genetics; D002923:Cilia / Q000235:genetics / Q000378:metabolism; D005124:Eye Abnormalities / Q000235:genetics* / Q000473:pathology; D016678:Genome; D006801:Humans; D009024:Morphogenesis / Q000235:genetics*; D009136:Muscular Dystrophies / Q000235:genetics / Q000473:pathology; D009154:Mutation; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics*; D010641:Phenotype; D011984:Sensory Receptor Cells / Q000378:metabolism* / Q000473:pathology; D058494:Walker-Warburg Syndrome / Q000235:genetics; D021601:trans-Golgi Network / Q000235:genetics
chemical_list: D017351:N-Acetylglucosaminyltransferases; C542575:B4GAT1 protein, human