EDGE CT: Genetic tools to study giant viruses

EDGE CT：研究巨型病毒的遗传工具

基本信息

批准号：
1922958
负责人：
Steven Wilhelm
金额：
$ 100.93万
依托单位：
University of Tennessee Knoxville
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2019
资助国家：
美国
起止时间：
2019-10-01 至 2024-09-30
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1922958&HistoricalAwards=false
关键词：
EDGE CT Genetic tools study

项目摘要

Non-Technical Abstract In recent years microbiologists have become aware of abundant "giant viruses" that infect protists. These viruses have also been implicated as components of the microbiomes of multicellular organisms. Many of the genes within these giants are considered novel for viruses: many genes encode for processes only associated with "living cellular organisms," like DNA replication or DNA repair, are found in these viruses. Studies suggest that these giant viruses have collected these genes from across all biology, leaving these viruses with genomes that can at best be described as mosaics. Moreover many of the hundreds of genes found in giant viruses remain completely new to science. In spite of this obvious wealth of information, we remain without tools to study how these giant virus genes function or if they are even necessary. This study will develop the tools needed to do genetic research in a model giant virus system (the Aureococcus anophagefferens Virus) and establish the ability of these tool sets to be extended to other giant viruses and their hosts.Technical AbstractWhile discovered only 15 years ago, giant viruses infecting protists have been shown to be distributed across ecosystems. These viruses contain functional genes encoding processes that can include light-dependent and independent DNA repair, sugar metabolism genes and genes associated with the process of translation. Phylogenetic examinations suggest these genes have been collected by rampant horizontal gene transfer. This study will develop protocols required to test gene function and activity in a giant virus (AaV) that infects the pelagophyte Aureococcus anophagefferens. A member of the Mimiviridae, AaV is a large virus with a complex complement of genes. The PIs and trainees will develop in vitro approaches to modify and transform AaV, including the introduction of antibiotic resistance and/or chromogenic/fluorogenic proteins for genetically tractable selections for use in subsequent gene replacement studies. The PIs will test protocols on other Mimiviridae and Phycodnaviridae that infect algae to determine tractability in other giant virus-host systems. Along with training of a postdoc, graduate and undergraduate students, this project will develop tools that allow for hypothesis driven testing in subsequent studies in a system where many ( 60%) of the gene products are annotated as hypothetical. The project will communicate protocols through training sessions as well as a dedicated workshop, and document all approaches in open access through protocols.io. Student journalists will produce material for public consumption, including short videos focusing on the positive role of viruses in ecology, the importance of technical reproducibility in science, and several on the fascinating nature of giant viruses.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.

近年来，非技术摘要微生物学家已经意识到感染了原生生物的丰富的“巨型病毒”。这些病毒也被认为是多细胞生物微生物的成分。这些巨人中的许多基因都被认为是新颖的病毒：许多与“活细胞生物”相关的过程编码的基因，例如在这些病毒中发现了DNA复制或DNA修复。研究表明，这些巨型病毒已从所有生物学中收集了这些基因，使这些病毒的基因组充其最多可以被描述为马赛克。此外，在巨型病毒中发现的数百个基因中的许多基因仍然是科学的全新基因。尽管有大量的信息，但我们仍然没有工具来研究这些巨型病毒基因的作用或甚至是必要的。这项研究将开发在模型巨型病毒系统（Aureoccus Anophagefferens病毒）中进行遗传研究所需的工具，并确定这些工具集将其扩展到其他巨型病毒及其宿主的能力。仅15年前才发现了巨大的病毒感染者在15年前发现的巨型病毒感染者，才显示出巨大的病毒感染者。这些病毒包含编码过程的功能基因，这些基因可以包括光依赖性和独立的DNA修复，糖代谢基因以及与翻译过程相关的基因。系统发育检查表明这些基因已通过猖ramp的水平基因转移收集。这项研究将开发在感染了骨植物巨卵球菌atophagefferens的巨型病毒（AAV）中测试基因功能和活性所需的方案。 AAV是Mimiviridae的成员，是一种大型病毒，具有复杂的基因补体。 PIS和受训者将开发体外方法来修饰和转化AAV，包括引入抗生素耐药性和/或发色/荧光蛋白，用于在后续基因替代研究中使用遗传学的选择。 PI将测试其他麦米维迪科和植物科动物的方案，这些方案感染藻类以确定其他巨型病毒宿主系统中的易干性。在培训博士后，研究生和本科生的培训之外，该项目将开发工具，在随后的研究中，在许多基因产物被注释为假设的系统中，在随后的研究中进行了假设驱动的测试。该项目将通过培训会议和专门的研讨会来传达协议，并记录通过协议的开放访问中的所有方法。学生记者将生产供公众消费的材料，包括关注病毒在生态学中的积极作用的简短视频，技术可重复性在科学中的重要性以及一些关于巨型病毒的迷人性质的重要性。该奖项反映了NSF的法定任务，并被认为是通过基金会的智力功能和广泛影响来评估CRETERIA的评估，认为这是值得的。