Inducible transgenes expressing EJC enriched mRNAs: a post-genomic innovation

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DESCRIPTION (provided by applicant): This revised proposal was originally submitted in response to PA-07-336, "Development of Animal Models and Related Biological Materials for Research." The research plan entails generation and initial characterization of murine transgenic models featuring novel inducible transgenes expressing the soluble, alternatively spliced form of human Tissue Factor (asTF), the principal trigger of coagulation whose aberrant expression and activity contributes to etiology of many debilitating disorders of the cardiovascular system, various forms of neoplasia, and proliferative diabetic retinopathy asTF expression will be restricted to the cells of monocyte/macrophage lineage, a well established source of bioactive TF in circulation. Innovative transgene design ensures that the produced asTF mRNA will be enriched in exon-exon junction complexes (EJC)-multimeric protein formations whose deposition on newly synthesized mRNA molecules was recently shown to be essential for effective mRNA nuclear export, intra-cytoplasmic targeting, translation and, most critically, post-translational modifications and export of secreted proteins. If successful, the project is very likely to substantially advance the field of transgenic animal research, as it will 1) establish and validate a conceptually novel approach to transgene design, via utilization of EJC enriched expression cassettes for optimal protein expression; 2) enhance the existing transgenic technologies of reversible in vivo gene expression in monocytes/macrophages, a cell type central to many vital (patho)physiologic processes in biology and disease, and 3) establish a technologically advanced in vivo approach to study circulating TF, whose function(s) in health and disease are of interest to many institutes within the NIH system, including NHLBI, NCI, NIDDK, NIA, and NEI. PUBLIC HEALTH RELEVANCE: This application proposes development of qualitatively novel inducible transgenes expressing mRNA enriched in exon-exon junction complexes (EJC)-multimeric protein formations whose deposition on newly synthesized mRNA molecules is essential for effective post-transcriptional mRNA processing. It entails generation of murine transgenic models expressing the alternatively spliced form of circulating human Tissue Factor (asTF), the principal trigger of coagulation whose elevated expression and activity is known to contribute to etiology of many debilitating disorders of the cardiovascular system, as well as various forms of neoplasia. If successful, the project will comprise a qualitative advance in transgenic technology as it will establish a principally novel methodology to study human TF, whose functions in health and disease fall within the categorical interests of several NIH institutes, most notably NHLBI, NIDDK, and NCI.