Structure/function studies of adenovirus capsid protein IX

病毒非常善于将其DNA传递到细胞中，这一观察结果已被用于开发高度减毒的病毒（不再引起疾病），这些病毒可以将任何感兴趣的基因传递到实验室中的细胞进行实验研究。 在本申请中，我们将继续我们的研究计划，致力于提高我们对Ad生物学基本方面的理解，这有助于其作为一种有效的基因递送系统的功能。 我们目前的工作集中在一种病毒蛋白质上，称为蛋白质IX。 这种蛋白质直接参与帮助病毒进入细胞，并且似乎还具有降低细胞对感染的天然防御的有效性的第二种功能。 我们将确定病毒和蛋白质如何能够做到这一点的机制，这将为细胞内的抗病毒蛋白以及病毒如何与它们相互作用提供新的见解。 最近，我们表明我们可以修饰蛋白IX，以使病毒感染靶向特定的细胞类型，如癌细胞。 然而，尽管递送是有效的，但这些病毒现在携带正常数量DNA的能力降低，原因尚不清楚。 我们将确定这种缺陷的性质，并探索纠正它的方法。总之，我们目前对pIX的工作将提供对病毒结构、感染和复制的深入了解，还将阐明作为抗病毒防御因子的细胞蛋白，进一步突出宿主和病原体之间正在进行的战斗。 此外，我们将改进我们生产基于Ad的基因递送系统的能力，这些系统可以靶向特定的细胞类型和组织，这最终将使许多利用Ad作为基因递送工具的研究人员受益。

Viruses are very good at delivering their DNA into cells, and this observation has been exploited to develop highly-attenuated viruses (no longer cause disease) that can deliver any gene-of-interest to cells in the laboratory for experimental study. In this application, we will continue our research program dedicated to improving our understanding of basic aspects of Ad biology that contributes to its function as an efficient gene delivery system. Our current work is focused on a viral protein, called protein IX. This protein is directly involved in helping the virus gain entry into the cell, and also appears to have a second function in reducing the effectiveness of the cell's natural defenses against infection. We will determine the mechanism of how the virus and protein are capable of doing this, which will provide novel insight into anti-viral proteins within the cell and how viruses interact with them. Recently, we showed that we could modify protein IX in order to target virus infection to specific cell types, such as cancer cells. However, although delivery was effective, these viruses now have a reduced ability to carry normal amounts of DNA, for reasons which are unclear. We will determine the nature of this deficiency and explore methods to correct it. Taken together, our current work on pIX will provide insight into virus structure, infection, and replication, and will also elucidate cellular proteins that act as anti-viral defense factors, further highlighting the ongoing battle between host and pathogen. In addition, we will refine our ability to produce Ad-based gene delivery systems that can be targeted to specific cell types and tissues, which ultimately will benefit many researchers utilizing Ad as a gene delivery tool.