<br>全基因组 CRISPR 筛选发现 TOB1 通过 IFN 和 EGFR 介导途径充当 FMDV 感染的关键宿主因子,PLoS Pathogens

当前位置： X-MOL 学术 › PLOS Pathog. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

全基因组 CRISPR 筛选发现 TOB1 通过 IFN 和 EGFR 介导途径充当 FMDV 感染的关键宿主因子
PLoS Pathogens ( IF 5.5 ) Pub Date : 2024-03-21 , DOI: 10.1371/journal.ppat.1012104
Gaochuang Peng _{1,

2} , Tianran Liu _{1,

2} , Xiaolan Qi ₂ , Yuzhe Wang ₁ , Jingjing Ren ₂ , Jiangling Peng ₂ , Xuguang Du ₁ , Siyu Hu ₁ , Sen Wu ₁ , Yaofeng Zhao ₁ , Dan Li ₂ , Haixue Zheng ₂

Affiliation

口蹄疫病毒（FMDV）与宿主的相互作用对于病毒感染极为重要，但对此的研究较少，不利于疫苗研发和口蹄疫控制。在这项研究中，我们设计了一个猪基因组规模的 CRISPR/Cas9 敲除文库，其中包含针对 16,886 个蛋白质编码基因的 93,859 个单向导 RNA、25 个长 ncRNA 和 463 个 microRNA。使用该文库，FMDV 感染所需的几个先前未报道的基因在 IBRS-2 细胞中在 FMDV 选择后高度富集。后续研究证实了 FMDV 对这些基因的依赖性，并且我们确定了 FMDV 相关宿主基因之一的功能作用： TOB1 （ERBB2.1的传感器）。 TOB1 -敲除通过正向调节RIG-I和MDA5的表达来显着抑制FMDV感染。我们进一步发现TOB1 -敲除导致转录因子CEBPA的mRNA转录本及其蛋白质的积累更多，这进一步增强了转录因子CEBPA的转录RIG-I和丙二醛5基因。此外， TOB1 -敲除可抑制 EGFR/ERBB2 途径介导的 FMDV 吸附和内化。最后，口蹄疫病毒的致命挑战TOB1 - 敲除小鼠证实删除TOB1抑制口蹄疫病毒感染体内。这些结果表明 TOB1 是猪口蹄疫病毒感染的关键宿主因子。

"点击查看英文标题和摘要"

A genome-wide CRISPR screening uncovers that TOB1 acts as a key host factor for FMDV infection via both IFN and EGFR mediated pathways

The interaction between foot-and-mouth disease virus (FMDV) and the host is extremely important for virus infection, but there are few researches on it, which is not conducive to vaccine development and FMD control. In this study, we designed a porcine genome-scale CRISPR/Cas9 knockout library containing 93,859 single guide RNAs targeting 16,886 protein-coding genes, 25 long ncRNAs, and 463 microRNAs. Using this library, several previously unreported genes required for FMDV infection are highly enriched post-FMDV selection in IBRS-2 cells. Follow-up studies confirmed the dependency of FMDV on these genes, and we identified a functional role for one of the FMDV-related host genes: TOB1 (Transducer of ERBB2.1). TOB1-knockout significantly inhibits FMDV infection by positively regulating the expression of RIG-I and MDA5. We further found that TOB1-knockout led to more accumulation of mRNA transcripts of transcription factor CEBPA, and thus its protein, which further enhanced transcription of RIG-I and MDA5 genes. In addition, TOB1-knockout was shown to inhibit FMDV adsorption and internalization mediated by EGFR/ERBB2 pathway. Finally, the FMDV lethal challenge on TOB1-knockout mice confirmed that the deletion of TOB1 inhibited FMDV infection in vivo. These results identify TOB1 as a key host factor involved in FMDV infection in pigs.

更新日期：2024-03-21

点击分享查看原文

点击收藏

公开下载

阅读更多本刊新发论文本刊介绍/投稿指南