AMPLIFICATION OF SELF PROCESSING RNA VECTORS IN PLANTS

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We propose a novel approach to large-scale production of custom RNA products using plant viral RNA vectors. This energy efficient, low cost and environmentally friendly method would generate long RNA molecules (not available through chemical synthesis) less expensively than alternative in vivo and in vitro transcription systems. the desired RNA fragment inserted in the laboratory into the RNA vector and amplified through the viral biological cycles could then, after only a few days' incubation in infected plants, be easily isolated as virion particles (which preclude RNA degradation and are an ideal storage form). after simple separation of chimeric viral RNA from the virion coat protein subunits, the desired RNA fragment would be cleaved out and separated from the rest of the viral RNA. For this purpose we suggest a novel cassette design where the desired RNA fragment within the RNA vector would be flanked by two latent ribozymes (inactive in vivo), which will not compromise the virus amplification. Their activation in vitro after isolation of the chimeric RNA will be accomplished through incubation in a solution environment not presented in vivo. PROPOSED COMMERCIAL APPLICATION: RNA is an important target for structural biology and biomedical studies including HIV and influenza viruses. It also has potential in antisense and ribozyme therapeutics, biotechnology products and molecular biology tools. Our technology would provide large amounts of custom RNAs less expensively and more efficiently than alternative methods. Our current business strategy is to license this new technology to larger companies such as Biosource Technologies.