Sunday, November 24, 2024
11/24/2024
DESCRIPTION (provided by applicant): In response to RFA DE-03-001 "Gene Discovery for Craniofacial Disorders" this application is for a unique program of efficient gene discovery in zebrafish to find candidates for human disease genes: Our specific aims are to discover zebrafish genes that when mutated result in (1) craniofacial bone dysgenesis, and (2) retinal degeneration. For both, a novel combination of powerful approaches toward gene discovery will be utilized. Lesions in genes will be introduced by viral-mediated insertional mutagenesis. Because the viral insertions serve to tag the mutated genes, the genes can be identified and cloned extremely rapidly, often within a few days. The mutants will be identified in highly efficient, directed, first generation (G1) screens, using embryos and larvae obtained by parthenogenesis and vitally stained with fluorescent dyes to reveal the phenotypes. Parthenogenesis and G1 screening eliminates months of time and saves rearing hundreds of thousands of fish required in the more usual forward-genetic strategies of raising families of mutants and screening F3 progeny. The new strains will be made rapidly available to other researchers using community resources, the Zebrafish Information Network to advertise the mutants and the Zebrafish International Resource Center to distribute them. Various favorable attributes of the zebrafish embryo and larva will permit detailed phenotypic studies and rapid characterization of the functions of the genes in facial bone and eye development.
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