Visualizing Influenza Viral RNA Packaging

 描述(申请人提供)：流感病毒构成公共卫生风险；在季节性流行期间，有效的疫苗和抗病毒药物减轻了疾病的严重性。然而，抗病毒耐药株和新型人畜共患流感株可以绕过这些古老的保护策略。了解有效地将流感基因组包装成子代病毒粒子所需的病毒和细胞特性将提供新的治疗靶点，并确定潜在的宿主范围限制因素。作为美国国立卫生研究院的博士后研究员，我开发了两种新的方法来可视化流感病毒RNA在生产性感染期间在活细胞中组装和运输的细胞内动态。这些方法将与传统的生化技术相结合，通过解决以下两个目标来阐明流感病毒RNA片段是如何包装的：目标1--确定所有八个病毒片段选择性地包装成子代病毒粒子的机制；以及目标2--确定介导流感病毒RNA从细胞核到质膜运输的宿主成分。拟议的工作将解决流感生物学中的许多悬而未决的问题，因为缺乏工具来可视化单个细胞内的多个病毒RNA片段，并跟踪病毒RNA在生产性感染期间在活细胞中的运动。这些研究将深入了解流感病毒如何在自然界中重新组合，并确定病毒RNA包装中涉及的新宿主因素，这些因素可作为潜在的治疗靶点。

 DESCRIPTION (provided by applicant): Influenza viruses pose a public health risk; effective vaccines and antivirals have alleviated the severity of disease during seasonal epidemics. However, antiviral resistance strains and novel zoonotic strains of influenza can bypass these age-old protection strategies. Understanding the viral and cellular properties required for efficient packaging of the influenza genome into progeny virions will provide novel therapeutic targets and define potential host-range restriction factors. As a post-doctoral fellow at the National Institutes of Health, I have developed two novel methods to visualize the intracellular dynamics of influenza viral RNA assembly and transport in live cells during a productive infection. These methodologies will be combined with traditional biochemical techniques to elucidate how influenza viral RNA segments are packaged by addressing the following two aims: Aim 1 - Determine the mechanism by which all eight viral RNA segments selectively package into progeny virions and Aim 2 - Identify host components that mediate transport of influenza viral RNA from the nucleus to the plasma membrane. The proposed work will address many outstanding questions in influenza biology that are unanswered because of a lack of tools to visualize multiple viral RNA segments within a single cell and to track viral RNA movement in live cells during a productive infection. These studies will provide insight into how influenza viruses reassort in nature and identify novel host factors involved in viral RNA packaging that can be pursued as potential therapeutic targets.