Effective inhibition of mRNA accumulation and protein expression of H5N1 avian influenza virus NS1 gene in vitro by small interfering RNAs
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Cell Line MeSH
- Virulence Factors antagonists & inhibitors genetics MeSH
- Real-Time Polymerase Chain Reaction MeSH
- Humans MeSH
- RNA, Small Interfering * MeSH
- Flow Cytometry MeSH
- Recombinant Fusion Proteins analysis genetics MeSH
- Genes, Reporter MeSH
- Artificial Gene Fusion MeSH
- Viral Nonstructural Proteins antagonists & inhibitors genetics MeSH
- Influenza A Virus, H5N1 Subtype genetics growth & development MeSH
- Blotting, Western MeSH
- Green Fluorescent Proteins analysis genetics MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- enhanced green fluorescent protein MeSH Browser
- Virulence Factors MeSH
- INS1 protein, influenza virus MeSH Browser
- RNA, Small Interfering * MeSH
- Recombinant Fusion Proteins MeSH
- Viral Nonstructural Proteins MeSH
- Green Fluorescent Proteins MeSH
Avian influenza has emerged as a devastating disease and may cross species barrier and adapt to a new host, causing enormous economic loss and great public health threats, and non-structural protein 1 (NS1) is a multifunctional non-structural protein of avian influenza virus (AIV) that counters cellular antiviral activities and is a virulence factor. RNA interference (RNAi) provides a powerful promising approach to inhibit viral infection specifically. To explore the possibility of using RNAi as a strategy against AIV infection, after the fusion protein expression plasmids pNS1-enhanced green fluorescent protein (EGFP), which contain the EGFP reporter gene and AIV NS1 as silencing target, were constructed and NS1-EGFP fusion protein expressing HEK293 cell lines were established, four small interfering RNAs (siRNAs) targeting NS1 gene were designed, synthesized, and used to transfect the stable cell lines. Flow cytometry, real-time quantitative polymerase chain reaction, and Western blot were performed to assess the expression level of NS1. The results suggested that sequence-dependent specific siRNAs effectively inhibited mRNA accumulation and protein expression of AIV NS1 in vitro. These findings provide useful information for the development of RNAi-based prophylaxis and therapy for AIV infection.
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