A CRISPR-Based Screen Identifies Genes Essential for West-Nile-Virus-Induced Cell Death.
A CRISPR-Based Screen Identifies Genes Essential for West-Nile-Virus-Induced Cell Death.
复制标题
DOI：
10.1016/j.celrep.2015.06.049
复制
发表时间：
2015-07-28
期刊：
影响因子：
8.8
通讯作者：
Wu H
中科院分区：
文献类型：
--
作者：
Ma H;Dang Y;Wu Y;Jia G;Anaya E;Zhang J;Abraham S;Choi JG;Shi G;Qi L;Manjunath N;Wu H
西尼罗河病毒（WNV）引起急性神经系统感染，伴随大量神经细胞死亡。然而，病毒诱导细胞死亡背后的机制知之甚少。使用含有77，406个靶向20，121个基因的sgRNA的文库，我们进行了全基因组筛选，然后用子文库进行了第二次筛选。在鉴定的基因中，EMC2、EMC3、SEL1L、DERL 2、UBE2G2、UBE2J1和HRD 1这七个基因因具有最强的表型而脱颖而出，其敲除在两种不同的WNV毒株和三种细胞系中赋予了针对WNV诱导的细胞死亡的强保护。有趣的是，敲除这些基因并不能阻断西尼罗河病毒的复制。因此，这些似乎是将WNV复制与下游细胞死亡途径联系起来的必需基因。此外，所有这些基因都属于内质网相关蛋白降解（ERAD）途径的事实表明，这可能是WNV诱导细胞死亡的主要驱动因素。
West Nile virus (WNV) causes an acute neurological infection attended by massive neuronal cell death. However, the mechanism(s) behind the virus-induced cell death is poorly understood. Using a library containing 77,406 sgRNAs targeting 20,121 genes, we performed a genome-wide screen followed by a second screen with a sub-library. Among the genes identified, seven genes, EMC2, EMC3, SEL1L, DERL2, UBE2G2, UBE2J1, and HRD1, stood out as having the strongest phenotype, whose knockout conferred strong protection against WNV-induced cell death with two different WNV strains and in three cell lines. Interestingly, knockout of these genes did not block WNV replication. Thus, these appear to be essential genes that link WNV replication to downstream cell death pathway(s). In addition, the fact that all of these genes belong to the endoplasmic reticulum-associated protein degradation (ERAD) pathway suggests that this might be the primary driver of WNV-induced cell death.