A Method for the Efficient Functional Examination of Mammalian Noncoding Sequences in Transgenic Zebrafish

sequence conservation is recognized as a reliable indicator of both coding and non-coding functional sequences. Consistent with this hypothesis, coding sequences may be readily identified based on evolutionary conservation. However, of the 5% of the human genome that is predicted to be functional based on conservation alone, less than a third actually encodes protein. The remaining conserved non-coding sequences probably regulate temporal, spatial, and quantitative aspects of gene expression. Consequently, there is significant interest in identifying and characterizing functional non-coding sequences. Inventors at Johns Hopkins University have developed a transposon-based transgenic strategy in zebrafish that facilitates systematic evaluation of non-coding sequences surrounding any locus in any vertebrate genome. This method relies on the integration of comparative genomic sequence analysis and transposon-based transgenesis in zebrafish. This techniques has already been applied to the analysis of both the zebrafish ret gene and its human ortholog (RET), identifying many regulatory sequences within and flanking these genes. Description (Set) Proposed Use (Set) 1. Identification of critical sequences of disease genes or disease gene candidates permitting their examination for disease-associated mutations 2. The evaluation of the functional impact of any identified candidate regulatory mutation 3. Generation of in vivo screening models for identification of chemicals modulating expression within or directly modulating the homeostasis/pathological status of identified lineages 4. Generation of in vivo screening models for mutagenesis strategies aimed at identification of genes with a critical role in the genesis or homeostasis of identified lineages.

Inventor(s): McCallion, Andrew Smythe

Type of Offer: Licensing

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