Small Interfering RNA Targeting M2 Gene Induces Effective and Long Term Inhibition of Influenza A Virus Replication 英文参考文献.docVIP

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Small Interfering RNA Targeting M2 Gene Induces Effective and Long Term Inhibition of Influenza A Virus Replication 英文参考文献.doc

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Small Interfering RNA Targeting M2 Gene Induces Effective and Long Term Inhibition of Influenza A Virus Replication 英文参考文献

SmallInterferingRNATargetingM2GeneInduces EffectiveandLongTermInhibitionofInfluenzaAVirus Replication Hong-YanSui1,Guang-YuZhao1,Jian-DongHuang2,Dong-YanJin2,Kwok-YungYuen1,Bo-JianZheng1* 1DepartmentofMicrobiology,TheUniversityofHongKong,Pokfulam,HongKongSAR,China,2DepartmentofBiochemistry,TheUniversityofHongKong,Pokfulam, HongKongSAR,China Abstract RNA interference (RNAi) provides a powerful new means to inhibit viral infection specifically. However, the selection of siRNA-resistantvirusesisamajorconcernintheuseofRNAiasantiviraltherapeutics.Inthisstudy,weconductedalentiviral vectorwithaH1-shorthairpinRNA(shRNA)expressioncassettetodeliversmallinterferingRNAs(siRNAs)intomammalian cells.Usingthisvectorthatalsoexpressesenhancedgreenfluorescenceprotein(EGFP)assurrogatemarker,stableshRNA- expressingcelllinesweresuccessfullyestablishedandtheinhibitionefficienciesofrationallydesignedsiRNAstargetingto conservedregionsofinfluenzaAvirusgenomewereassessed.TheresultsshowedthatasiRNAtargetinginfluenzaM2gene (siM2) potently inhibited viral replication. The siM2 was not only effective for H1N1 virus but also for highly pathogenic avian influenza virus H5N1. In addition to its M2 inhibition, the siM2 also inhibited NP mRNA accumulation and protein expression.AlongterminhibitioneffectofthesiM2wasdemonstratedandtheemergenceofsiRNA-resistantmutantsin influenza quasispecies wasnotobserved. Takentogether,our study suggested that M2gene mightbeanoptimal RNAi targetforantiviraltherapy.ThesefindingsprovideusefulinformationforthedevelopmentofRNAi-basedprophylaxisand therapyforhumaninfluenzavirusinfection. Citation:SuiH-Y,ZhaoG-Y,HuangJ-D,JinD-Y,YuenK-Y,etal.(2009)SmallInterferingRNATargetingM2GeneInducesEffectiveandLongTermInhibitionof InfluenzaAVirusReplication.PLoSONE4(5):e5671.doi:10.1371/journal.pone.0005671 Editor:DingXiangLiu,InstituteofMolecularandCellBiology,Singapore ReceivedFebruary18,2009;AcceptedMay3,2009;PublishedMay22,2009 Copyright:?2009Suietal.Thisisanopen-accessarticledi