鉴定Ⅱ型伪狂犬病毒感染细胞中的功能长链非编码 RNA。

Identification of functional lncRNAs in pseudorabies virus type II infected cells.

机构信息

MOA Key Laboratory of Animal Virology, Department of Veterinary Sciences, Zhejiang University, Hangzhou 310058, China; Institute of Animal Preventive Sciences, Department of Veterinary Sciences, Zhejiang University, Hangzhou 310058, China; Collaborative Innovation Center and State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University, Hangzhou 310058, China.

出版信息

Vet Microbiol. 2020 Mar;242:108564. doi: 10.1016/j.vetmic.2019.108564. Epub 2019 Dec 23.

DOI:10.1016/j.vetmic.2019.108564

PMID:32122616

Abstract

Long noncoding RNAs (lncRNAs) play important roles in the antiviral responses. However, little is known about the identification and functions of swine lncRNAs in response to pseudorabies virus type II (PRV-II). Here, we detected the expression profiles of host lncRNAs from a wild-type (PRV-II DX) and gE/TK deficient (gE-TK-PRV) PRV-II infected cells. RNA-seq identified 664 differentially expressed (DE) lncRNAs from PRV-DX infected cells, 654 DE lncRNAs from gE-TK-PRV infected cells and 276 DE lncRNAs between PRV-DX and gE-TK-PRV infected cells. The potential functions of the significant differentially expressed (SDE) lncRNAs were involved in interleukin secretion, axon extension and metabolic process based on the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Moreover, the expression patterns of sixteen SDE lncRNAs determined by RT-qPCR exhibited high correlation (r > 0.95) with those by RNA-seq results. Western blotting assay displayed the lncA02830 did not code for protein, and the silencing of lncA02830 could significantly up-regulate the transcription levels of IRF3, IFNβ as well as MX1 and inhibit the replication of PRV-II. Taken together, these data highlighted the potentials of lncRNA as targets for antiviral therapy and provided some novel knowledge of the mechanisms underlying the host interaction with PRV-II.

摘要

长链非编码 RNA（lncRNA）在抗病毒反应中发挥重要作用。然而，对于猪 lncRNA 在应对伪狂犬病病毒 II 型（PRV-II）中的鉴定和功能知之甚少。在这里，我们检测了野生型（PRV-II DX）和 gE/TK 缺失（gE-TK-PRV）PRV-II 感染细胞中宿主 lncRNA 的表达谱。RNA-seq 从 PRV-DX 感染的细胞中鉴定出 664 个差异表达（DE）lncRNA，从 gE-TK-PRV 感染的细胞中鉴定出 654 个 DE lncRNA，从 PRV-DX 和 gE-TK-PRV 感染的细胞中鉴定出 276 个 DE lncRNA。基于基因本体论（GO）和京都基因与基因组百科全书（KEGG）数据库，显著差异表达（SDE）lncRNA 的潜在功能涉及白细胞介素分泌、轴突延伸和代谢过程。此外，通过 RT-qPCR 确定的十六个 SDE lncRNA 的表达模式与 RNA-seq 结果高度相关（r>0.95）。Western blotting 检测显示 lncA02830 不编码蛋白质，lncA02830 的沉默可显著上调 IRF3、IFNβ 以及 MX1 的转录水平，并抑制 PRV-II 的复制。综上所述，这些数据强调了 lncRNA 作为抗病毒治疗靶点的潜力，并为宿主与 PRV-II 相互作用的机制提供了一些新的认识。