EAGER: Toward a tractable genetic model of DNA virus - Drosophila interaction

DNA病毒是卫生保健中的一个难题，也是一种广泛使用的有害生物种群管理工具，特别是在农业中。然而，研究人员缺乏良好的模型来研究DNA病毒与其宿主之间的相互作用。本项目旨在创建一个可操作的遗传模型，用于研究宿主/DNA病毒相互作用，使用最广泛使用的模式生物之一果蝇。这一进展将允许通过标准实验室技术对病毒进行遗传操作，以更好地了解毒力的分子遗传基础、宿主操作以及传染病的出现和演变。研究人员还将优化裸病毒在细胞系中的生长，使未来的实验成为可能。此外，该项目将从一个转校生项目中培训本科研究人员，为他们提供动手科学训练，并有助于扩大科学参与。第一个目标是创造新的细胞系并优化病毒复制的条件。迄今为止，可用的细胞系已被用于培养病毒，但收效甚微。利用基于自然宿主组织和优化条件的新细胞系有望改善病毒在细胞系中的复制。第二个目标是将大的双链DNA基因组克隆到细菌人工染色体中，以便于生产，然后表明使用标准的定点诱变和基于pcr的重组技术对病毒进行遗传操作是简单的。项目完成后，研究人员将开发出一种多功能、易于处理的系统，用于研究宿主/DNA病毒的共同进化，可用于指导DNA病毒疾病的治疗和生物防治策略。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

DNA viruses are a difficult problem in health care and a widely-used tool for managing pest populations, particularly in agriculture. However, researchers lack good models to study the interactions between DNA viruses and their hosts. This project aims to create a tractable genetic model for studying host/DNA virus interaction using fruit flies (Drosophila), one of the most widely used model organisms. This advance would allow for genetic manipulation of the virus through standard laboratory techniques to better understand the molecular genetic basis of virulence, host manipulation, and the emergence and evolution of infectious disease. Researchers will also optimize the growth of the nudivirus in cell lines, making possible a drove of experiments in the future. In addition, the project would train undergraduate researchers from a transfer student program to provide them with hands-on scientific training and contribute to broadening participation in science. The first objective is to create new cell lines and optimize conditions for viral replication. To date available cell lines have been used to grow the virus with limited success. Utilizing new cell lines based on natural host tissues and optimized conditions promises to improve viral replication in cell lines. The second objective is to clone the large, double stranded DNA genome into a bacterial artificial chromosome for ease of production, then show that genetic manipulation of the virus is straightforward using standard site-directed mutagenesis and PCR-based recombinant techniques. Upon project completion, researchers will have developed a versatile, tractable system for studying host/DNA virus coevolution that can be used to inform treatment of DNA virus diseases and biocontrol strategies.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.