title: Rapid identification of kidney cyst mutations by whole exome sequencing in zebrafish.
abstract: Forward genetic approaches in zebrafish have provided invaluable information about developmental processes. However, the relative difficulty of mapping and isolating mutations has limited the number of new genetic screens. Recent improvements in the annotation of the zebrafish genome coupled to a reduction in sequencing costs prompted the development of whole genome and RNA sequencing approaches for gene discovery. Here we describe a whole exome sequencing (WES) approach that allows rapid and cost-effective identification of mutations. We used our WES methodology to isolate four mutations that cause kidney cysts; we identified novel alleles in two ciliary genes as well as two novel mutants. The WES approach described here does not require specialized infrastructure or training and is therefore widely accessible. This methodology should thus help facilitate genetic screens and expedite the identification of mutants that can inform basic biological processes and the causality of genetic disorders in humans.
authors.affiliation: Department of Cell Biology, Duke University, 333 B, Nanaline Duke Building, Durham, NC 27710, USA.
authors.forename: Sean
authors.initials: S
authors.lastname: Ryan
authors.forename: Jason
authors.initials: J
authors.lastname: Willer
authors.forename: Lindsay
authors.initials: L
authors.lastname: Marjoram
authors.forename: Jennifer
authors.initials: J
authors.lastname: Bagwell
authors.forename: Jamie
authors.initials: J
authors.lastname: Mankiewicz
authors.forename: Ignaty
authors.initials: I
authors.lastname: Leshchiner
authors.forename: Wolfram
authors.initials: W
authors.lastname: Goessling
authors.forename: Michel
authors.initials: M
authors.lastname: Bagnat
authors.forename: Nicholas
authors.initials: N
authors.lastname: Katsanis
mesh_terms: D000818:Animals; D003560:Cysts / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D059472:Exome / Q000235:genetics*; D008040:Genetic Linkage; D007668:Kidney / Q000473:pathology*; D018613:Microscopy, Confocal; D016296:Mutagenesis / Q000235:genetics*; D009841:Oligonucleotides / Q000235:genetics; D015027:Zebrafish / Q000235:genetics*
chemical_list: D009841:Oligonucleotides
keywords: Cilia; Exome sequencing; Kidney cyst