Real-time resolution of point mutations that cause phenovariance in mice

• DOI: 10.1073/pnas.1423216112
• Publication Date: 2015-01-21T06:19:22Z
• Authors: Tao Wang; Xiaowei Zhan; Chun-Hui Bu; Stephen Lyon; David Pratt; Sara Hildebrand; Jin Huk Choi; Zhao Zhang; Ming Zeng; Kuan-wen Wang; Emre Turer; Zhe Chen; Duanwu Zhang; Tao Yue; Ying Wang; Hexin Shi; Jianhui Wang; Lei Sun; Jeff SoRelle; William McAlpine; Noelle Hutchins; Xiaoming Zhan; Maggy Fina; Rochelle Gobert; Jiexia Quan; McKensie Kreutzer; Stephanie Arnett; Kimberly Hawkins; Ashley Leach; Christopher Tate; Chad Daniel; Carlos Reyna; Lauren Prince; Sheila Davis; Joel Purrington; Rick Bearden; Jennifer Weatherly; Danielle White; Jamie Russell; Qihua Sun; Miao Tang; Xiaohong Li; Lindsay Scott; Eva Marie Y. Moresco; Gerald M. McInerney; Gunilla B. Karlss...; Yang Xie; Bruce Beutler
• Keywords: Male; 0303 health sciences; Genetic Linkage; Quantitative Trait Loci; Pedigree; Mice; 03 medical and health sciences; Phenotype; Animals; Point Mutation; Female; Genes, Lethal; Alleles

With the wide availability of massively parallel sequencing technologies, genetic mapping has become rate limiting step in mammalian forward genetics. Here we introduce a method for real-time identification N-ethyl-N-nitrosourea-induced mutations that cause phenotypes mice. All are identified by whole exome G1 progenitor and their zygosity is established G2/G3 mice before phenotypic assessment. Quantitative qualitative traits, including lethal effects, single or multiple combined pedigrees then analyzed with Linkage Analyzer, software program detects significant linkage between individual aberrant scores presents processed data as Manhattan plots. As alleles genes acquired through mutagenesis, pooled "superpedigrees" created to analyze effects. Our distinguished from conventional methods because it permits (1) unbiased declaration mappable phenotypes, those incompletely penetrant (2), automated causative concurrent screening, without need outcross mutant another strain backcross them, (3) exclusion not involved interest. We validated our approach Analyzer 47 45 previously known adaptive immune phenotypes; analysis also implicated 474 associated function. The described here mice, limited only rates production screening.