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

项目摘要 SARS-CoV-2是全球COVID-19大流行的罪魁祸首。除了急性症状，如 呼吸衰竭和其他流感样症状，这种病毒最近已经牵连到许多慢性 症状，统称为COVID-19急性后后遗症（PASC，也称为“长 COVID“）。PASC的许多症状与神经系统有关，如认知功能障碍、脑雾、 记忆力减退、感觉问题、抑郁、焦虑和失眠。与我们所理解的 SARS-CoV-2引起COVID-19的严重急性症状，该病毒引起PACS的机制是 没有很好地理解。为了方便研究COVID-19和PASC，我们一直在生成三种类型的 果蝇基因试剂。第一个是一组转基因果蝇， 以细胞类型或时间特异性方式条件表达单一病毒蛋白。第二个是一系列 转基因苍蝇允许有条件地表达已发现在物理上 与SARS-CoV-2蛋白相互作用。第三个是一个多功能基因陷阱系列的集合， 研究果蝇基因的功能丧失表型和表达模式， SARS-CoV-2人类相互作用者。科学家们可以将这三种类型的试剂联合收割机进行复杂的 研究病毒和人类蛋白质是否在活的生物体中功能性地相互作用，或者 果蝇和人的同源基因在体内具有相同的分子功能。在本研究中， 我们建议进一步扩大研究范围，以完善馆藏。此外，我们将在体内进行 表征迄今为止产生的试剂，以提供表型和基因表达信息， 促进这些试剂在研究界的使用。这些基因工具已经或将公开提供 通过果蝇布卢明顿库存中心，收集的信息将记录在FlyBase中。 这种转基因和突变果蝇的收集将极大地促进编码蛋白质的体内表征 在SARS-CoV-2基因组及其相互作用物中，因此，它将为抗击COVID-19提供有价值的信息- 19、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.