Treatment with ribavirin and alpha interferon inhibited replicon activity, suggesting that both act on the level of RNA replication, transcription, or ...ArticlePDFAvailableRepliconSystemforLassaVirusJanuary2005JournalofVirology78(24):13793-803DOI:10.1128/JVI.78.24.13793-13803.2004SourcePubMedAuthors:MeikeHassBernhardNochtInstituteforTropicalMedicineUtaGölnitzStefanieChristineBeckerUniversityofVeterinaryMedicineHannoverBeateBecker-ZiajaRobertKochInstitutShowall5authorsHideDownloadfull-textPDFReadfull-textDownloadfull-textPDFReadfull-textDownloadcitationCopylinkLinkcopiedReadfull-textDownloadcitationCopylinkLinkcopiedCitations(105)References(61)Figures(7)AbstractandFiguresLassavirusisendemictoWestAfricaandcauseshemorrhagicfeverinhumans.Tofacilitatethefunctionalanalysisofthisvirus,arepliconsystemwasdevelopedbasedonLassavirusstrainAV.Genomicandantigenomicminigenomes(MG)wereconstructedconsistingoftheintergenicregionofSRNAandareportergene(Renillaluciferase)inantisenseorientation,flankedbythe5'and3'untranslatedregionsofSRNA.MGswereexpressedunderthecontroloftheT7promoter.Nucleoprotein(NP),Lprotein,andZproteinwereexpressedfromplasmidscontainingtheT7promoterandinternalribosomalentrysite.TransfectionofcellsstablyexpressingT7RNApolymerase(BSRT7/5)withMGintheformofDNAorRNAandplasmidsfortheexpressionofNPandLproteinresultedinhighlevelsofRenillaluciferaseexpression.Therepliconsystemwasoptimizedwithrespecttotheratioofthetransfectedconstructsandbymodifyingthe5'endoftheMG.Maximumactivitywasobserved24to36haftertransfectionwithasignal-to-noiseratioof2to3logunits.NorthernblotanalysisprovidedevidenceforreplicationandtranscriptionoftheMG.Zproteindownregulatedrepliconactivityclosetobackgroundlevels.Treatmentwithribavirinandalphainterferoninhibitedrepliconactivity,suggestingthatbothactonthelevelofRNAreplication,transcription,orribonucleoproteinassembly.Inconclusion,thisstudydescribesthefirstrepliconsystemforahighlypathogenicarenavirus.Itisatoolforinvestigatingthemechanismsofreplicationandtr