title: Sleeping Beauty Transposon Mutagenesis as a Tool for Gene Discovery in the NOD Mouse Model of Type 1 Diabetes.
abstract: A number of different strategies have been used to identify genes for which genetic variation contributes to type 1 diabetes (T1D) pathogenesis. Genetic studies in humans have identified >40 loci that affect the risk for developing T1D, but the underlying causative alleles are often difficult to pinpoint or have subtle biological effects. A complementary strategy to identifying "natural" alleles in the human population is to engineer "artificial" alleles within inbred mouse strains and determine their effect on T1D incidence. We describe the use of the Sleeping Beauty (SB) transposon mutagenesis system in the nonobese diabetic (NOD) mouse strain, which harbors a genetic background predisposed to developing T1D. Mutagenesis in this system is random, but a green fluorescent protein (GFP)-polyA gene trap within the SB transposon enables early detection of mice harboring transposon-disrupted genes. The SB transposon also acts as a molecular tag to, without additional breeding, efficiently identify mutated genes and prioritize mutant mice for further characterization. We show here that the SB transposon is functional in NOD mice and can produce a null allele in a novel candidate gene that increases diabetes incidence. We propose that SB transposon mutagenesis could be used as a complementary strategy to traditional methods to help identify genes that, when disrupted, affect T1D pathogenesis.
authors.affiliation: Immunology and Diabetes Unit, St. Vincent's Institute, Fitzroy, Victoria 3065, Australia.
authors.forename: Colleen M
authors.initials: CM
authors.lastname: Elso
authors.affiliation: Immunology and Diabetes Unit, St. Vincent's Institute, Fitzroy, Victoria 3065, Australia Department of Medicine, University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: Edward P F
authors.initials: EP
authors.lastname: Chu
authors.affiliation: Immunology and Diabetes Unit, St. Vincent's Institute, Fitzroy, Victoria 3065, Australia Department of Medicine, University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: May A
authors.initials: MA
authors.lastname: Alsayb
authors.affiliation: Immunology and Diabetes Unit, St. Vincent's Institute, Fitzroy, Victoria 3065, Australia.
authors.forename: Leanne
authors.initials: L
authors.lastname: Mackin
authors.affiliation: Immunology and Diabetes Unit, St. Vincent's Institute, Fitzroy, Victoria 3065, Australia.
authors.forename: Sean T
authors.initials: ST
authors.lastname: Ivory
authors.affiliation: Immunology and Diabetes Unit, St. Vincent's Institute, Fitzroy, Victoria 3065, Australia Department of Medicine, University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: Michelle P
authors.initials: MP
authors.lastname: Ashton
authors.affiliation: Research School of Biology, The Australian National University, Canberra, ACT 2601, Australia.
authors.forename: Stefan
authors.initials: S
authors.lastname: Bröer
authors.affiliation: Dendritic Cell Research, ANZAC Research Institute, Concord, New South Wales 2139, Australia.
authors.forename: Pablo A
authors.initials: PA
authors.lastname: Silveira
authors.affiliation: Immunology and Diabetes Unit, St. Vincent's Institute, Fitzroy, Victoria 3065, Australia Department of Medicine, University of Melbourne, Parkville, Victoria 3010, Australia tbrodnicki@svi.edu.au.
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Brodnicki
mesh_terms: D000818:Animals; D056905:Chromosome Breakpoints; D004251:DNA Transposable Elements; D003922:Diabetes Mellitus, Type 1 / Q000235:genetics*; D004195:Disease Models, Animal; D005260:Female; D018628:Gene Dosage; D015870:Gene Expression; D017930:Genes, Reporter; D056726:Genetic Association Studies; D005822:Genetic Vectors / Q000235:genetics*; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D008822:Mice, Transgenic; D016254:Mutagenesis, Insertional; D009154:Mutation; D040641:Quantitative Trait Loci
chemical_list: D004251:DNA Transposable Elements
keywords: Serinc1; Slc16a10; amino acid transporter; forward genetics; reverse genetics