Molecular dissection studies of mammalian reovirus proteins using reverse genetics.

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Mammalian reovirus has played an important role in our understanding of important biological mechanisms: virus entry, synthesis of nucleic acids and proteins, virus evolution, and more. In addition to its interest as a fundamental research tool, possible applications are now envisaged for this fascinating virus. In fact, some viruses, such as reovirus, could become our allies rather than just enemies to fight. Its ability to infect human beings, as well as a large number of animal species, without causing disease in humans, is a further incentive to develop “virotherapeutic” agents from this virus. For example, it is now well established that reovirus preferentially infects and kills cancer cells. It can be further envisaged developing the virus as a gene-transfer vector. On the contrary, new pathogenic isolates of reovirus have been reported in the last few years, suggesting the possibility of emerging reoviruses that could represent new threats to humans and animals. During the last 25 years, members of the laboratory have significantly contributed to our understanding of the virus. Molecular biology, biochemistry, and classical genetics approaches, have been especially useful to dissect the structure and function of many viral proteins. In 2007, we witnessed a major progress with the development of a new and powerful method of plasmid-based reverse genetics which could be used to produce various viral mutants, using genetic engineering. This was a major progress, now allowing to study the effect of various changes to the viral proteins in the context of the viral replicative cycle. We were among the few laboratories to readily adopt this promising approach. It has been used extensively in the last few years to produce and study new reovirus variants that possess potentially interesting properties. In the studies proposed herein, three different viral capsid proteins will be studied: these play different, yet incompletely understood, roles, especially in virus entry, synthesis of viral nucleic acids and control of the host-cell response to infection. In addition to the net gains in fundamental knowledge, our research program is motivated by longer-term objectives aimed at developing and optimizing applications of the virus as well as achieving better control of the possible emerging pathogens.