International Mammalian Genome Society

logo18th International Mouse Genome Conference

17-22 October 2004, Seattle, USA


POSTER 66 - MAPPING AND POSITIONAL CLONING OF ENU-INDUCED MOUSE DEVELOPMENTAL MUTANTS WITH A WHOLE GENOME SNP PANEL AND EXON-BASED SEQUENCING

Moran JL 1, Yun Y 1, Brown A 1, Bjork BC 1, Tran P 1, Li C 2, Salinger AP 3, Poirier C 3, Zhang B 1, Montgomery K 1, Bishop C 3, Justice MJ 3, Wiltshire T 4, Kwiatkowski DJ 1, Kucherlapati R 1, Beier DR 1

1 Brigham and Women's Hospital, Harvard Medical School, Boston, MA, United States, 2 Dana Farber Cancer Institute, Harvard School of Public Health, Boston, MA, United States, 3 Baylor College of Medicine, Houston, TX, United States, 4 Genomics Institute of the Novartis Research Foundation, San Diego, CA, United States

ENU mutagenesis has proven to be an efficient means for generating novel developmental mutations in the mouse. However, the task of localizing and characterizing these mutations remains a significant barrier to their general utilization. We have established a consortium for the genetic mapping, phenotypic characterization and positional cloning of ENU-induced mutations affecting developmental processes.  In order to expeditiously map mutations at moderate resolution, we have assembled a genome wide panel of over 400 single nucleotide polymorphisms (SNPs).  Thus far, confirmed linkage of 5 recessive mutants has been obtained by genotyping 10-11 affected mice.  Genetic intervals of 21-38 Mb identified with the SNP panel were subsequently reduced to 2-13 Mb by genotyping additional affecteds with microsatellite markers.  The SNP panel offers several advantages over conventional microsatellite marker panels: higher marker density, which enables the detection of whole genome linkage information, and the ability to map mutations with small numbers of mice independent of genetic background. Preliminary chromosomal localization of a genetic modifier suggests that the panel potentially has use beyond mapping ENU-induced mutations.

In order to identify causative mutations in ENU mutants, a high-throughput pipeline for sequencing candidate genes in recombinant intervals has been developed.  The pipeline is exon-based and includes semi-automated primer design, primer ordering, amplification, sequencing, and data analysis.  Both the SNP genotyping and sequencing platforms are available to interested investigators.  Using these genetic mapping and sequencing methods, we have identified a causative mutation in the mummy mutant, a recessive perinatal lethal mutant with an epidermal permeability barrier defect.

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