Department of Microbiology, The University of Chicago, Chicago, IL 60637, USA.
Center for Research Informatics, The University of Chicago, Chicago, IL 60637, USA.
Cell Rep. 2018 Apr 10;23(2):596-607. doi: 10.1016/j.celrep.2018.03.045.
The emergence of influenza A viruses (IAVs) from zoonotic reservoirs poses a great threat to human health. As seasonal vaccines are ineffective against zoonotic strains, and newly transmitted viruses can quickly acquire drug resistance, there remains a need for host-directed therapeutics against IAVs. Here, we performed a genome-scale CRISPR/Cas9 knockout screen in human lung epithelial cells with a human isolate of an avian H5N1 strain. Several genes involved in sialic acid biosynthesis and related glycosylation pathways were highly enriched post-H5N1 selection, including SLC35A1, a sialic acid transporter essential for IAV receptor expression and thus viral entry. Importantly, we have identified capicua (CIC) as a negative regulator of cell-intrinsic immunity, as loss of CIC resulted in heightened antiviral responses and restricted replication of multiple viruses. Therefore, our study demonstrates that the CRISPR/Cas9 system can be utilized for the discovery of host factors critical for the replication of intracellular pathogens.
甲型流感病毒(IAV)从动物宿主中出现,对人类健康构成了巨大威胁。由于季节性疫苗对动物源病毒株无效,而新传播的病毒可能很快产生耐药性,因此仍需要针对 IAV 的宿主定向治疗方法。在这里,我们在人类肺上皮细胞中进行了全基因组 CRISPR/Cas9 敲除筛选,使用的是一株人源 H5N1 禽流感病毒分离株。在 H5N1 选择后,几个参与唾液酸生物合成和相关糖基化途径的基因被高度富集,包括 SLC35A1,这是一种对 IAV 受体表达和病毒进入至关重要的唾液酸转运体。重要的是,我们已经确定 capicua(CIC)是细胞内固有免疫的负调节剂,因为 CIC 的缺失会导致抗病毒反应增强,并限制多种病毒的复制。因此,我们的研究表明,CRISPR/Cas9 系统可用于发现对细胞内病原体复制至关重要的宿主因子。