title: Efficient generation and mapping of recessive developmental mutations using ENU mutagenesis.
abstract: Treatment with N-ethyl-N-nitrosourea (ENU) efficiently generates single-nucleotide mutations in mice. Along with the renewed interest in this approach, much attention has been given recently to large screens with broad aims; however, more finely focused studies have proven very productive as well. Here we show how mutagenesis together with genetic mapping can facilitate the rapid characterization of recessive loci required for normal embryonic development. We screened third-generation progeny of mutagenized mice at embryonic day (E) 18.5 for abnormalities of organogenesis. We ascertained 15 monogenic mutations in the 54 families that were comprehensively analyzed. We carried out the experiment as an outcross, which facilitated the genetic mapping of the mutations by haplotype analysis. We mapped seven of the mutations and identified the affected locus in two lines. Using a hierarchical approach, it is possible to maximize the efficiency of this analysis so that it can be carried out easily with modest infrastructure and resources.
authors.affiliation: Genetics Division, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Bruce J
authors.initials: BJ
authors.lastname: Herron
authors.forename: Weining
authors.initials: W
authors.lastname: Lu
authors.forename: Cherie
authors.initials: C
authors.lastname: Rao
authors.forename: Shanming
authors.initials: S
authors.lastname: Liu
authors.forename: Heiko
authors.initials: H
authors.lastname: Peters
authors.forename: Roderick T
authors.initials: RT
authors.lastname: Bronson
authors.forename: Monica J
authors.initials: MJ
authors.lastname: Justice
authors.forename: J David
authors.initials: JD
authors.lastname: McDonald
authors.forename: David R
authors.initials: DR
authors.lastname: Beier
mesh_terms: D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D004247:DNA / Q000235:genetics; D004252:DNA Mutational Analysis; D005314:Embryonic and Fetal Development / Q000235:genetics*; D005038:Ethylnitrosourea / Q000633:toxicity*; D005260:Female; D005808:Genes, Recessive; D006239:Haplotypes; D008297:Male; D051379:Mice; D008805:Mice, Inbred A; D008807:Mice, Inbred BALB C; D016296:Mutagenesis; D009154:Mutation; D010641:Phenotype; D011247:Pregnancy
chemical_list: D004247:DNA; D005038:Ethylnitrosourea