KSEF R&D Excellence: Development of a Novel Biotechnology Tool Based on RNA Recombination

Expression of beneficiary genes in selected plants promises major advancement in agriculture. The goal of this project is to develop a completely novel biotechnology tool that can be useful to complement 'conventional' recombinant DNA technologies. This new technology will be based on RNA recombination, a process that is extensively studied in the PI's lab. RNA recombination by plant viruses is a powerful process, since .itutilizes already existing and functional viral genetic elements, such as genes and replication signals. Based on the PI's and others' results, it is now possible to promote RNA recombination in plants using plant RNA viruses. The PI terms this novel approach as "recombinant RNA technology" . Recombinant RNA technology win be used to generate and express cytotoxic genes in the infected cens in order to induce resistance against plant viruses. The idea is to use RNA viruses of plants, such as tombusviruses, and their recombination machinery to create functional messenger (m)RNAs from nonfunctional mRNAs that carry a cytotoxic gene by recombining them with viral sequences in a targeted (site-specific) fashion at the RNA level in plants. The recombinant mRNAs generated in the cytoplasm of infected cells would then be able to express cytotoxic proteins only in the infected cens leading to rapid cell death. The rapid cell death is expected to cause extreme resistance against viruses, as we demonstrated in our preliminary experiments. The major advantage of this technology over the traditional DNA-based technology is that the recombinant protein win not be synthesized in uninfected cells/plants. Overall, this research may lead to the development of a significantly safer tool for future disease management than those currently available. Application of the technology should be beneficial in preventing future major losses from virus infections, which are presently difficult to controL