Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, Missouri, USA
Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, Missouri, USA.
J Virol. 2018 Dec 10;93(1). doi: 10.1128/JVI.01324-18. Print 2019 Jan 1.
Noroviruses (NoVs) are a leading cause of gastroenteritis worldwide, yet host factors that restrict NoV replication are not well understood. Here, we use a CRISPR activation genome-wide screening to identify host genes that can inhibit murine norovirus (MNoV) replication in human cells. Our screens identified with high confidence 49 genes that can inhibit MNoV infection when overexpressed. A significant number of these genes are in interferon and immune regulation signaling networks, but surprisingly, the majority of the genes identified are neither associated with innate or adaptive immunity nor associated with any antiviral activity. Confirmatory studies of eight of the genes validate the initial screening data. Mechanistic studies on demonstrated a conserved role of the molecule in mouse and human cells in restricting MNoV in a step of infection after viral entry. Furthermore, we demonstrate that two isoforms of have differential antiviral activity. Taken together, these data provide a resource for understanding norovirus biology and demonstrate a robust methodology for identifying new antiviral molecules. Norovirus is one of the leading causes of food-borne illness worldwide. Despite its prevalence, our understanding of norovirus biology is limited due to the difficulty in growing human norovirus and a lack of an animal model. Murine norovirus (MNoV) is a model norovirus system because MNoV replicates robustly in cell culture and in mice. To identify host genes that can restrict norovirus replication when overexpressed, we performed genome-wide CRISPR activation screens to induce gene overexpression at the native locus through recruitment of transcriptional activators to individual gene promoters. We found 49 genes that could block murine norovirus replication in human cells. Several of these genes are associated with classical immune signaling pathways, while many of the molecules we identified have not been previously associated with antiviral activity. Our data are a resource for those studying noroviruses, and we provide a robust approach to identify novel antiviral genes.
诺如病毒（NoV）是全球范围内导致肠胃炎的主要原因，但宿主限制 NoV 复制的因素尚未得到很好的理解。在这里，我们使用 CRISPR 激活全基因组筛选来鉴定可以在人类细胞中抑制鼠诺如病毒（MNoV）复制的宿主基因。我们的筛选以高置信度鉴定了 49 个在过表达时可以抑制 MNoV 感染的基因。这些基因中有相当数量存在于干扰素和免疫调节信号网络中，但令人惊讶的是，大多数鉴定的基因既与先天免疫或适应性免疫无关，也与任何抗病毒活性无关。对其中 8 个基因的验证性研究验证了初始筛选数据。对 进行的机制研究表明，该分子在感染后病毒进入的一个感染步骤中在小鼠和人类细胞中具有保守的限制 MNoV 的作用。此外，我们证明了 有两种同工型，具有不同的抗病毒活性。总之，这些数据为理解诺如病毒生物学提供了资源，并展示了一种用于鉴定新抗病毒分子的强大方法。诺如病毒是全球食源性疾病的主要原因之一。尽管它很普遍，但由于难以培养人类诺如病毒和缺乏动物模型，我们对诺如病毒生物学的理解有限。鼠诺如病毒（MNoV）是一种模型诺如病毒系统，因为 MNoV 在细胞培养物和小鼠中能强烈复制。为了鉴定在过表达时可以限制诺如病毒复制的宿主基因，我们进行了全基因组 CRISPR 激活筛选，通过将转录激活因子募集到单个基因启动子上来在天然基因座上诱导基因过表达。我们发现了 49 个可以阻断人类细胞中 MNoV 复制的基因。其中一些基因与经典免疫信号通路有关，而我们鉴定的许多分子以前与抗病毒活性无关。我们的数据为研究诺如病毒的人提供了资源，并且为鉴定新的抗病毒基因提供了一种强大的方法。
