Researcher Chen Jilong from the Institute of Microbiology, Chinese Academy of Sciences/Fujian Agriculture and Forestry University has long been engaged in research on the pathogenesis of influenza virus, antiviral immunology, mechanism of tumor infection induced by viral infection, and tumor immunology. Recently, its laboratory used Arraystar Human LncRNA microarray to discover a novel long-chain non-coding RNA that regulates host antiviral response, NRAV, which regulates antiviral responses by inhibiting interferon-stimulated gene transcription, in antiviral innate immunity. The reaction plays an important regulatory role. The research results were published in the international top journal Cell, Cell, Host & Microbe (influence factor 12.194). (Chip experiment provided by Kang Cheng technical service) Research Background: The flu is an important zoonotic infectious disease that is highly infectious and contagious. After the influenza virus infects the host, the viral RNA is recognized by the host's pathogen recognition receptor, activates the antiviral natural immune response, and inhibits viral replication. LncRNA has been a hot research field in the biological and medical fields in the past five years. It has been reported that lncRNA plays an important role in many cellular processes, but the role of lncRNA in the antiviral natural immune response is still unclear. Professor Chen Jilong's research group aims to use the Arraystar Human LncRNA chip to systematically study the function of lncRNA in influenza virus replication and host antiviral natural immune response. Research ideas: In order to systematically detect the effect of virus on host lncRNA, the authors first used the American Arraystar Human LncRNA chip to detect the expression of lncRNA in influenza A virus (IAV) infected and non-infected alveolar epithelial cells (3 cases each). Among them, the expression level of lncRNA-NRAV was significantly decreased in IAV-infected cells, and the non-coding features and full-length sequences of NRAV were confirmed by means of bioinformatics analysis and molecular biological means. The in vitro and in vivo experiments of cells and transgenic mice showed that NRAV can significantly promote the replication and virulence of IAV, indicating that NRAV is a negative regulator of the antiviral natural immune response. The authors further investigated the mechanism by which NRAV promotes IAV replication, overexpressing NRAV in virus-infected cells, and detecting gene expression changes by whole-genome expression profiling. GO analysis revealed that many differentially expressed genes are involved in pathogen infection and viral replication, and that gene expression of many interferon-stimulated genes has changed significantly. The authors studied the mechanisms by which NRAV affects interferon-stimulated gene expression at multiple levels. ChIP-PCR experiments revealed that NRAV significantly altered the level of histone modification in the promoter regions of key interferon-stimulated genes IFITM3 and MxA, thereby inhibiting gene transcription of IFITM3 and MxA. Further, it was also found by RNA pull-down and RIP experiments that NRAV specifically binds to the multifunctional transcription factor ZONAB and interacts with each other, thereby affecting the transcriptional regulation of MxA. Technical route: The results show: A. The clustering map showed differential expression of lncRNA, significant difference between lncRNA and RT-PCR validation; Significance: This study reveals important regulatory functions of lncRNA in antiviral natural immune responses. In virus-infected cells, the expression level of long-chain non-coding RNA-NRAV is significantly reduced. In vitro and in vivo experiments have shown that NRAV can significantly promote the replication of IAV, as a negative regulator affects the antiviral natural immune response. This study identified a new host antiviral response lncRNA that promotes the interaction between the virus and the host. It has important scientific significance for revealing the pathogenesis of acute infectious viruses such as influenza, and is a new antiviral drug. Drug targets are provided. Original source: NRAV, a Long Noncoding RNA, Modulates Antiviral Responses through Suppression of Interferon-Stimulated Gene Transcription. Cell Host & Microbe, 2014, 16(5): 616-626. Automatic Air Hose Reel,Auto Rewind Air Hose Reel,Self Retracting Air Hose Reel,Air Compressor Retractable Hose Reel NINGBO QIKAI ENVIRONMENTAL TECHNOLOGY CO.,LTD , https://www.water-hose-reel.com
B. The clustering map shows differentially expressed mRNA, interferon-stimulated gene clustering map;
C. GO analysis of differentially expressed genes screened by whole genome expression profiling;
D. ChIP-PCR experiments showed that NRAV altered histone modifications of MxA and IFITM3.