Reverse genetics study of mammalian reovirus proteins: fundamental studies and possible applications

哺乳动物呼肠病毒在我们理解重要的生物学机制方面发挥了重要作用：病毒进入、核酸和蛋白质合成、病毒进化等等。除了将其作为基础研究工具之外，还设想了可能的应用。一些病毒，如呼肠孤病毒，可能成为我们的盟友，而不仅仅是我们的敌人。呼肠孤病毒感染人类和大范围动物物种的能力表明开发新应用的可能性。例如，现在已经确定呼肠孤病毒优先感染并杀死癌细胞；因此，它是抗癌“病毒疗法”的一个有吸引力的候选者。在过去20年左右的时间里，实验室的成员一直非常活跃地研究呼肠孤病毒复制的各个方面。分子生物学、生物化学和经典遗传学方法对于解剖许多病毒蛋白的结构和功能特别有用。建议继续进行这项工作，以便更好地了解病毒复制及其对感染细胞的影响，这对未来治疗应用的发展至关重要。自2007年以来，一种被称为“反向遗传学”的新方法允许引入呼肠孤病毒基因组的各种变化。生产各种病毒突变体变得相对容易，不仅依赖于随机突变，而且使用基因工程和“定点”诱变。这是研究病毒复制方面的一个重大进展，否则无法进行研究。将研究三种不同的蛋白质：它们在病毒进入、感染不同细胞的能力、在这些细胞中复制和杀死这些细胞方面发挥着不同的鲜为人知的作用。这些研究将有助于实现开发和优化呼肠孤病毒在动物和人类生物学中的应用的长期目标。

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 much more. In addition to its interest as a fundamental research tool, possible applications are envisaged. Some viruses, such as reovirus, could become our allies rather than just enemies to fight. The ability of reovirus to infect humans and a large range of animal species suggests the possibility of developing novel applications. For example, it is now well established that reovirus preferentially infects and kills cancer cells; it is thus an attractive candidate for "virotherapy" against cancer. In the last 20 years or so, members of the laboratory have been very active in the study of various aspects of reovirus replication. Molecular biology, biochemistry, and classical genetics approaches, have been especially useful to dissect the structure and function of many viral proteins. It is proposed to pursue this work in order to develop a better understanding of viral replication and its effects on infected cells, an aspect judged essential for the future development of therapeutic applications. Since 2007, a novel method known as "reverse genetics" allows introduction of various changes to the reovirus genome. It becomes relatively easy to produce various viral mutants, relying not only to random mutations but using genetic engineering and "site-directed" mutagenesis. This is a major progress to study aspects of viral replication that could not be studied otherwise. Three different proteins will be studied: these play different poorly known roles in virus entry, ability to infect different cells, replicate in and kill these cells. These studies will contribute to long-term objectives aimed at developing and optimizing reovirus' applications in animal and human biology.