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Simulated RNA Interference in the T4 Bacteriophage Lytic Cycle

Greenlee, Jacob
Abstract
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RNA interference pathways have shown promise in disrupting the translation of protein products in eukaryotic cells and organisms. These studies have used siRNA or miRNA, but not mRNA. Bacteria lack the genes to produce the RNAi enzymes found in eukaryotes. In this study, an attempt to simulate an RNAi pathway in hopes of using E. coli as a model organism for mRNA viral therapeutics was made. Part of the T4 K10 bacteriophage T-holin gene (~125kb) has been successfully cloned into the pET expression vector. To disrupt viral mRNA and translation, a complimentary clone needs to be generated and has yet to be accomplished. Initial controls have demonstrated that the presence of the plasmid and pseudogene expression of the partial T-holin mRNA do not interfere with viral replication.
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University of Wyoming Libraries