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

哺乳动物呼肠孤病毒蛋白的反向遗传学研究：基础研究和可能的应用

• 批准号: 155580-2012
• 负责人: Lemay, Guy
• 金额: $ 1.89万
• 依托单位: Université de Montréal
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2012
• 资助国家: 加拿大
• 起止时间: 2012-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=505688
• 关键词: Reverse; genetics; study; mammalian; reovirus

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.

哺乳动物呼肠孤病毒在我们理解重要的生物学机制方面发挥了重要作用：病毒入侵、核酸和蛋白质的合成、病毒进化等。除了它作为基础研究工具的兴趣之外，还设想了可能的应用。一些病毒，如呼肠孤病毒，可能会成为我们的盟友，而不仅仅是敌人。呼肠孤病毒能够感染人类和大范围的动物物种，这表明有可能开发新的应用。例如，现在已经确定呼肠孤病毒优先感染和杀死癌细胞；因此，它是治疗癌症的“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，实验室成员一直非常积极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剖析许多病毒蛋白质的结构和功能特别有用。建议继续这项工作，以便更好地了解病毒复制及其对受感染细胞的影响，这一方面被认为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”的新方法允许在呼肠孤病毒基因组中引入各种变化。制造各种病毒突变变得相对容易，不仅依靠随机突变，而且使用基因工程和“定点”突变。这是在研究病毒复制方面的重大进展，否则无法研究这些方面。将研究三种不同的蛋白质：它们在病毒进入、感染不同细胞、在这些细胞内复制和杀死这些细胞的能力中扮演着不同的鲜为人知的角色。这些研究将有助于实现旨在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。