International Mammalian Genome Society

logo18th International Mouse Genome Conference

17-22 October 2004, Seattle, USA


ORAL PRESENTATION

WEDNESDAY OCTOBER 20

12.00pm – 12.15pm

A SENSITIZED MOUSE MUTAGENESIS SCREEN FOR NOVEL GENES REGULATING NEURAL CREST CELL CEVELOPMENT

Watkins-Chow DE, Silver DL, Matera I, Baxter LL, Elliott G, Rivas C, Incao A, Pavan WJ

National Human Genome Institute, NIH, Bethesda, MD, United States

We have established an enhancer screen in mouse to identify novel mutations disrupting development of the neural crest derived melanocyte lineage. Our focused phenotype screen is based on the prediction that ENU induced mutations acting synergistically with the neural crest transcription factor, Sox10, will cause white spotting larger than the typical spot (1.3 +/- 1.0 % of total ventral body area) that we observe in Sox10LacZ/+ heterozygous mutant control animals. In screening 100 first generation pedigrees, we have identified 2 heritable phenotypes that display a significant increase in white spotting on the Sox10LacZ/+ heterozygous background. The BALB/cJ and C57BL/6J mixed genetic background of our cross has facilitated subsequent mapping of these ENU induced mutations to two independent regions of the genome distinct from loci previously implicated in known coat color mutants. Positional cloning is currently underway to identify the mutations responsible for these ENU induced alleles. In addition to the second generation phenotype screen, third generation embryos from 20 lines have been screened for both dominant and recessive embryonic phenotypes that alter expression of the Sox10LacZ reporter gene. Work is underway to confirm heritability of a phenotype selected in this embryonic screen that displays a disorganized pattern of gene expression in the region of the developing dorsal root ganglia. DNA samples from cryopreserved first generation mice provide an invaluable resource for recovering live animals from any identified phenotype and for additional sequence-based, non-phenotype driven approaches to mutation detection.

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