title: Inhibition of influenza A virus matrix and nonstructural gene expression using RNA interference.
abstract: Influenza antiviral drugs that use protein inhibitors can lose their efficacy as resistant strains emerge. As an alternative strategy, we investigated the use of small interfering RNA molecules (siRNAs) by characterizing three siRNAs (M747, M776 and M832) targeting the influenza matrix 2 gene and three (NS570, NS595 and NS615) targeting the nonstructural protein 1 and 2 genes. We also re-examined two previously reported siRNAs, M331 and M950, which target the matrix 1 and 2 genes. Treatment with M331-, M776-, M832-, and M950-siRNAs attenuated influenza titer. M776-siRNA treated cells had 29.8% less infectious virus than cells treated with the previously characterized siRNA, M950. NS570-, NS595- and NS615-siRNAs reduced nonstructural protein 1 and 2 expression and enhanced type I interferon expression by 50%. Combination siRNA treatment attenuated 20.9% more infectious virus than single siRNA treatment. Our results suggest a potential use for these siRNAs as an effective anti-influenza virus therapy.
authors.affiliation: Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, Morgantown, WV, USA; Department of Microbiology, Immunology and Cell Biology, School of Medicine, West Virginia University, Morgantown, WV, United States.
authors.forename: Cynthia M
authors.initials: CM
authors.lastname: McMillen
authors.affiliation: Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, Morgantown, WV, USA; Department of Microbiology, Immunology and Cell Biology, School of Medicine, West Virginia University, Morgantown, WV, United States.
authors.forename: Donald H
authors.initials: DH
authors.lastname: Beezhold
authors.affiliation: Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, Morgantown, WV, USA.
authors.forename: Francoise M
authors.initials: FM
authors.lastname: Blachere
authors.affiliation: Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, Morgantown, WV, USA.
authors.forename: Sreekumar
authors.initials: S
authors.lastname: Othumpangat
authors.affiliation: Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, Morgantown, WV, USA.
authors.forename: Michael L
authors.initials: ML
authors.lastname: Kashon
authors.affiliation: Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, Morgantown, WV, USA; Department of Microbiology, Immunology and Cell Biology, School of Medicine, West Virginia University, Morgantown, WV, United States. Electronic address: jnoti@cdc.gov.
authors.forename: John D
authors.initials: JD
authors.lastname: Noti
mesh_terms: D000818:Animals; D002460:Cell Line; D004285:Dogs; D015967:Gene Expression Regulation, Viral; D023061:Gene Order; D006801:Humans; D009980:Influenza A virus / Q000235:genetics* / Q000502:physiology; D007251:Influenza, Human / Q000235:genetics / Q000378:metabolism / Q000821:virology; D007370:Interferon Type I / Q000096:biosynthesis; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D014763:Viral Matrix Proteins / Q000235:genetics* / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000378:metabolism; D010948:Viral Plaque Assay; D014779:Virus Replication
chemical_list: D007370:Interferon Type I; C488936:M1 protein, Influenza A virus; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D014763:Viral Matrix Proteins; D017361:Viral Nonstructural Proteins
keywords: Antivirals; Gene expression; Influenza virus; Matrix protein 1 (M1); Matrix protein 2 (M2); Nonstructural protein 1 (NS1); Nonstructural protein 2 (NS2); RNA interference (RNAi); Small interfering RNA (siRNA)