title: Genome-wide ENU mutagenesis in combination with high density SNP analysis and exome sequencing provides rapid identification of novel mouse models of developmental disease.
abstract: BACKGROUND
  Mice harbouring gene mutations that cause phenotypic abnormalities during organogenesis are invaluable tools for linking gene function to normal development and human disorders. To generate mouse models harbouring novel alleles that are involved in organogenesis we conducted a phenotype-driven, genome-wide mutagenesis screen in mice using the mutagen N-ethyl-N-nitrosourea (ENU).
  
  
  RESULTS
  ENU was injected into male C57BL/6 mice and the mutations transmitted through the germ-line. ENU-induced mutations were bred to homozygosity and G3 embryos screened at embryonic day (E) 13.5 and E18.5 for abnormalities in limb and craniofacial structures, skin, blood, vasculature, lungs, gut, kidneys, ureters and gonads. From 52 pedigrees screened 15 were detected with anomalies in one or more of the structures/organs screened. Using single nucleotide polymorphism (SNP)-based linkage analysis in conjunction with candidate gene or next-generation sequencing (NGS) we identified novel recessive alleles for Fras1, Ift140 and Lig1.
  
  
  CONCLUSIONS
  In this study we have generated mouse models in which the anomalies closely mimic those seen in human disorders. The association between novel mutant alleles and phenotypes will lead to a better understanding of gene function in normal development and establish how their dysfunction causes human anomalies and disease.
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, Melbourne, Australia. georgina.caruana@monash.edu
authors.forename: Georgina
authors.initials: G
authors.lastname: Caruana
authors.forename: Peter G
authors.initials: PG
authors.lastname: Farlie
authors.forename: Adam H
authors.initials: AH
authors.lastname: Hart
authors.forename: Stefan
authors.initials: S
authors.lastname: Bagheri-Fam
authors.forename: Megan J
authors.initials: MJ
authors.lastname: Wallace
authors.forename: Michael S
authors.initials: MS
authors.lastname: Dobbie
authors.forename: Christopher T
authors.initials: CT
authors.lastname: Gordon
authors.forename: Kerry A
authors.initials: KA
authors.lastname: Miller
authors.forename: Belinda
authors.initials: B
authors.lastname: Whittle
authors.forename: Helen E
authors.initials: HE
authors.lastname: Abud
authors.forename: Ruth M
authors.initials: RM
authors.lastname: Arkell
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Cole
authors.forename: Vincent R
authors.initials: VR
authors.lastname: Harley
authors.forename: Ian M
authors.initials: IM
authors.lastname: Smyth
authors.forename: John F
authors.initials: JF
authors.lastname: Bertram
mesh_terms: D000483:Alleles; D000818:Animals; D000013:Congenital Abnormalities / Q000196:embryology / Q000209:etiology / Q000235:genetics*; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000235:genetics; D004195:Disease Models, Animal; D005038:Ethylnitrosourea / Q000633:toxicity*; D059472:Exome / Q000187:drug effects*; D016326:Extracellular Matrix Proteins / Q000235:genetics; D005260:Female; D055106:Genome-Wide Association Study; D005838:Genotype; D018095:Germ-Line Mutation / Q000187:drug effects; D059014:High-Throughput Nucleotide Sequencing; D006720:Homozygote; D055458:Left-Right Determination Factors / Q000235:genetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL / Q000235:genetics*; D016296:Mutagenesis; D009153:Mutagens / Q000633:toxicity*; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide
chemical_list: D016326:Extracellular Matrix Proteins; C482018:Fras1 protein, mouse; C000607892:LIG1 protein, human; D055458:Left-Right Determination Factors; C000607894:Lig1 protein, mouse; D009153:Mutagens; D011088:DNA Ligases; D000072481:DNA Ligase ATP; D005038:Ethylnitrosourea