STTR Phase I: Scaleable, Inexpensive Production of siRNA in E. coli

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This Small Business Technology Transfer Phase I project develops molecular biology, biochemical engineering, and purification techniques to provide "small interfering RNA" or siRNA molecules on at large-scale and at low cost compared to available techniques. In recent years, siRNA's have been recognized as potent gene-silencing agents via "RNA interference" (RNAi). Genes for important diseases such as cancer, HIV, and hepatitis have already been shown to be controllable by siRNA action. Applications of siRNA's now warrant large-scale manufacture of siRNAs for the purposes of RNAi associated studies and as pharmaceutical agents. Unique to the technology is our well demonstrated evidence that RNA insert sequences can be harbored in larger ribosomal RNA (rRNA) carriers and produced at high levels within E. coli by conventional fermentation. Following fermentation, the desired siRNA is excised and purified for large-scale production of the RNA product.The broader impacts of this research is to provide a general, large-scale production tool applicable to a wide range of future novel RNA products. In addition to siRNA production, the methods proposed here can be used for large-scale production of micro-RNAs, aptamers, antisense therapeutics, or other short RNA molecules that are also of considerable interest for structural and pharmacological applications. The associated products and RNA expression systems are expected to have considerable commercial value, first in the research community and eventually as pharmaceuticals. This project will also result in fundamental scientific knowledge regarding RNA structure, enzymatic action, and purification of particular RNA molecules.