C636 Aedes albopictus Cells Have a Dysfunctional Antiviral RNA Interference Response 英文参考文献.docVIP

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C636 Aedes albopictus Cells Have a Dysfunctional Antiviral RNA Interference Response 英文参考文献.doc

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C636 Aedes albopictus Cells Have a Dysfunctional Antiviral RNA Interference Response 英文参考文献

C6/36AedesalbopictusCellsHaveaDysfunctional AntiviralRNAInterferenceResponse DougE.Brackney1*,JaclynC.Scott2,FumihikoSagawa2,JimmyE.Woodward3,NeilA.Miller3,FayeD. Schilkey3,JoannMudge3,JeffreyWilusz2,KenE.Olson2,CarolD.Blair2,GregoryD.Ebel1 1Department of Pathology, University of New Mexico School of Medicine, Albuquerque, New Mexico, United States of America, 2Arthropod-Borne and Infectious Diseases Laboratory, Department ofMicrobiology, Immunology and Pathology, ColoradoStateUniversity, FortCollins,Colorado,United States ofAmerica, 3National CenterforGenomeResources,SantaFe,NewMexico,UnitedStatesofAmerica Abstract MosquitoesrelyonRNAinterference(RNAi)astheirprimarydefenseagainstviralinfections.Tothisend,thecombinationof RNAiandinvertebratecellculturesystemshasbecomeaninvaluabletoolinstudyingvirus-vectorinteractions.Nevertheless,a recentstudyfailedtodetectanactiveRNAiresponsetoWestNilevirus(WNV)infectioninC6/36(Aedesalbopictus)cells,a mosquitocelllinefrequentlyusedtostudyarthropod-borneviruses(arboviruses).Therefore,wesoughttodetermineifWNV activelyevadesthehost’sRNAiresponseorifC6/36cellshaveadysfunctionalRNAipathway.C6/36andDrosophilamelanogaster S2cellswereinfectedwithWNV(Flaviviridae),Sindbisvirus(SINV,Togaviridae)andLaCrossevirus(LACV,Bunyaviridae)andtotal RNArecoveredfromcelllysates.SmallRNA(sRNA)librarieswereconstructedandsubjectedtohigh-throughputsequencing.In S2cells,virus-derivedsmallinterferingRNAs(viRNAs)fromallthreeviruseswerepredominantly21ntinlength,ahallmarkofthe RNAipathway.However,inC6/36cells,viRNAswereprimarily17ntinlengthfromWNVinfectedcellsand26–27ntinlengthin SINVandLACVinfectedcells.Furthermore,theorigin(positiveornegativeviralstrand)anddistribution(positionalongviral genome)ofS2cellgeneratedviRNApopulationswasconsistentwithpreviouslypublishedstudies,buttheprofileofsRNAs isolatedfromC6/36cellswasaltered.Intotal,theseresultssuggestthatC6/36cellslackafunctionalantiviralRNAiresponse. Thesefindingsareanalogoustothetype-Iinterferondeficiencydes