title: Inhibition of Marburg virus protein expression and viral release by RNA interference.
abstract: High mortality rates and lack of an available vaccine against Marburg haemorrhagic fever (MHF) highlight the need for a defensive therapy against MHF and greater knowledge of the causative agent, the Marburg virus (MARV). Here, RNA interference (RNAi) is employed to destroy MARV transcripts, disrupting replication and allowing analysis of various roles of MARV proteins. Small interfering RNAs (siRNAs) homologous to three MARV transcripts (NP, VP35 and VP30) were co-transfected into cells with plasmids encoding the corresponding nucleocapsid proteins. The resulting decrease in MARV nucleocapsid-protein levels was shown to be specific, as siRNA that was not homologous to the MARV genome did not decrease the levels of viral nucleocapsid proteins. Additionally, transcript levels of double-stranded RNA (dsRNA)-sensor proteins, the dsRNA-activated protein kinase and 2',5'-oligoadenylate synthetase 1 remained unchanged, suggesting that the decrease in viral proteins was not a result of activation of the antiviral properties of the interferon system. Subsequently, siRNAs were shown to reduce intracellular viral proteins in MARV-infected cells and viral material released into the medium. Targeted reduction of VP30 downregulated the intracellular levels of all other viral proteins, suggesting that VP30 plays an essential role for transcription/replication. The efficient reduction of MARV replication also suggests that RNAi may provide an agent against MHF.
authors.affiliation: Max Planck Institute for Infection Biology, Schumannstrasse 21/22, Campus Charité Mitte, 10117 Berlin, Germany.
authors.forename: Trent
authors.initials: T
authors.lastname: Fowler
authors.affiliation: Institute of Virology, Philipps University Marburg, Robert-Koch-Strasse 17, 35037 Marburg, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Bamberg
authors.affiliation: Institute of Virology, Philipps University Marburg, Robert-Koch-Strasse 17, 35037 Marburg, Germany.
authors.forename: Peggy
authors.initials: P
authors.lastname: Möller
authors.affiliation: Institute of Virology, Philipps University Marburg, Robert-Koch-Strasse 17, 35037 Marburg, Germany.
authors.forename: Hans-Dieter
authors.initials: HD
authors.lastname: Klenk
authors.affiliation: Max Planck Institute for Infection Biology, Schumannstrasse 21/22, Campus Charité Mitte, 10117 Berlin, Germany.
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Meyer
authors.affiliation: Institute of Virology, Philipps University Marburg, Robert-Koch-Strasse 17, 35037 Marburg, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Becker
authors.affiliation: Max Planck Institute for Infection Biology, Schumannstrasse 21/22, Campus Charité Mitte, 10117 Berlin, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Rudel
mesh_terms: D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D015967:Gene Expression Regulation, Viral; D006367:HeLa Cells; D006801:Humans; D029024:Marburgvirus / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014709:Vero Cells; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000378:metabolism*
chemical_list: D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D034741:RNA, Small Interfering; C094001:VP30 protein, Marburg virus; D014758:Viral Core Proteins; D014764:Viral Proteins