a Oklahoma Center for Respiratory and Infectious Diseases , Oklahoma State University , Stillwater , OK , USA.
b The Lundberg-Kienlen Lung Biology and Toxicology Laboratory, Department of Physiological Sciences , Oklahoma State University , Stillwater , OK , USA.
RNA Biol. 2019 Mar;16(3):340-353. doi: 10.1080/15476286.2019.1572448. Epub 2019 Jan 28.
Long non-coding RNAs (lncRNAs) are a new arm of gene regulatory mechanism as discovered by sequencing techniques and follow-up functional studies. There are only few studies on lncRNAs as related to gene expression regulation and anti-viral activity during influenza virus infection. We sought to identify and characterize lncRNAs involved in influenza virus replication. Using RNA sequencing analysis, we found that 1,912 lncRNAs were significantly changed in human lung epithelial A549 cells infected with influenza A/Puerto Rico/8/34. Gene ontology analysis on neighboring genes of these lncRNAs revealed that the genes involved in type I interferon signaling and cellular response were highly enriched. Seven selected up-regulated lncRNAs (AC015849.2, RP-1-7H24.1, PSMB8-AS1, CTD-2639E6.9, PSOR1C3, AC007283.5 and RP11-670E13.5) were verified by real-time PCR. These lncRNAs were also induced by other two influenza H1N1 virus strains (A/WSN/1933 and A/Oklahoma/3052/09) and interferon β1. Repression of PSMB8 antisense RNA 1 (PSMB8-AS1) using CRISPR interference reduced viral mRNA and protein levels as well as the release of progeny influenza virus particles. Our study suggests that lncRNA PSMB8-AS1 could be a new host factor target for developing antiviral therapy against influenza virus infection.
长链非编码 RNA（lncRNA）是通过测序技术和后续功能研究发现的基因调控机制的一个新分支。在流感病毒感染过程中，关于 lncRNA 与基因表达调控和抗病毒活性的相关研究还很少。我们试图鉴定和描述参与流感病毒复制的 lncRNA。通过 RNA 测序分析，我们发现感染甲型流感病毒 A/Puerto Rico/8/34 后，人肺上皮细胞 A549 中有 1912 个 lncRNA 显著变化。对这些 lncRNA 邻近基因进行基因本体论分析显示，涉及 I 型干扰素信号和细胞反应的基因高度富集。通过实时 PCR 验证了 7 个上调的 lncRNA（AC015849.2、RP-1-7H24.1、PSMB8-AS1、CTD-2639E6.9、PSOR1C3、AC007283.5 和 RP11-670E13.5）。另外两种流感 H1N1 病毒株（A/WSN/1933 和 A/Oklahoma/3052/09）和干扰素 β1 也能诱导这些 lncRNA。利用 CRISPR 干扰抑制 PSMB8 反义 RNA 1（PSMB8-AS1）可降低病毒 mRNA 和蛋白水平以及子代流感病毒颗粒的释放。我们的研究表明，lncRNA PSMB8-AS1 可能是针对流感病毒感染开发抗病毒治疗的新宿主因子靶标。
