Expansion and characterization of the Drosophila Toolkit to study SARS-CoV-2

项目摘要 SARS-CoV-2是全球新冠肺炎大流行的罪魁祸首。除了急性症状，如 呼吸衰竭和其他类似流感的症状，这种病毒最近被牵连到许多慢性 症状，统称为新冠肺炎急性后遗症(PASC，也称为 CoVID‘)。PASC的许多症状与神经系统有关，如认知功能障碍、脑雾、 记忆力丧失、感觉问题、抑郁、焦虑和失眠。与我们对如何 SARS-CoV-2会导致严重的新冠肺炎急性症状，这种病毒引起PACs的机制是 不是很清楚。为了促进新冠肺炎和PASC的研究，我们已经生成了三种类型的 果蝇中的遗传试剂。第一个是转基因果蝇的集合，它可以让 以细胞类型或时间特定的方式有条件地表达单个病毒蛋白。第二个是一系列 转基因果蝇，允许有条件地表达物理上已发现的单个人类蛋白质 与SARS-CoV-2蛋白之一相互作用。第三种是一组多功能的基因陷阱系，它们允许一种 为了研究苍蝇基因的功能丧失表型和表达模式， SARS-CoV-2人类传播者。科学家可以将这三种试剂结合起来，进行复杂的 研究病毒和人类蛋白质是否在活的有机体中进行功能性相互作用的实验，或者是否 同源基因在蝇和人体内的功能具有相同的分子功能。在这项研究中， 我们建议进一步扩大我们的研究工具包，以完成收集。此外，我们还将在体内进行表演 迄今产生的提供表型和基因表达信息的试剂的表征 促进这些试剂在研究界的使用。这些遗传工具已经或将公开提供。 通过果蝇布鲁明顿种群中心，收集的信息将被记录在FlyBase中。 这种转基因和突变果蝇的收集将极大地促进体内编码蛋白质的表征 在SARS-CoV-2基因组及其相互作用因子中。因此，它将为抗击COVID提供有价值的信息- 19和PASC。

Project Summary SARS-CoV-2 is responsible for the global COVID-19 pandemic. In addition to acute symptoms such as respiratory failure and other flu-like symptoms, this virus has been more recently implicated in numerous chronic symptoms, which are collectively referred to as Post-Acute Sequelae of COVID-19 (PASC, also known as ‘long COVID’). Many symptoms of PASC are related to the nervous system, such as cognitive dysfunction, brain fog, memory loss, sensory problems, depression, anxiety and insomnia. Compared to our understanding on how SARS-CoV-2 causes severe acute symptoms of COVID-19, the mechanism by which this virus causes PACS is not well understood. To facilitate the study of COVID-19 and PASC, we have been generating three types of genetic reagents in the fruit fly Drosophila melanogaster. The first is a collection of transgenic flies that allows conditional expression of single viral proteins in a cell-type or time specific manner. The second is a series of transgenic flies that allows conditional expression of individual human proteins that have been found to physically interact with one of the SARS-CoV-2 proteins. The third is a collection of versatile gene-trap lines that allow one to investigate the loss-of-function phenotype and expression pattern of the fly gene that correspond to one of the SARS-CoV-2 human interactors. Scientists can combine these three types of reagents to perform sophisticated experiments to study whether the viral and human proteins functionally interact in a living organism, or whether the function of the homologous genes in fly and human share the same molecular function in vivo. In this study, we propose to further expand our research toolkit to complete the collection. In addition, we will perform in vivo characterization of the reagents generated so far to provide phenotypic and gene expression information to facilitate the use of these reagents in the research community. These genetic tools are or will be publicly available through the Drosophila Bloomington Stock Center and the information collected will be documented in FlyBase. This collection of transgenic and mutant flies will greatly facilitate the in vivo characterization of proteins encoded in the SARS-CoV-2 genome and its interactors,. It will therefore provide valuable information to combat COVID- 19 and PASC.