Broad-Spectrum Antiviral Activity of RNA Interference against Four Genotypes of Japanese Encephalitis Virus Based on Single MicroRNA Polycistrons 英文参考文献.docVIP

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Broad-Spectrum Antiviral Activity of RNA Interference against Four Genotypes of Japanese Encephalitis Virus Based on Single MicroRNA Polycistrons 英文参考文献.doc

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Broad-Spectrum Antiviral Activity of RNA Interference against Four Genotypes of Japanese Encephalitis Virus Based on Single MicroRNA Polycistrons 英文参考文献

Broad-SpectrumAntiviralActivityofRNAInterference againstFourGenotypesofJapaneseEncephalitisVirus BasedonSingleMicroRNAPolycistrons ZhiqiangWu.,YingXue.,BeiWang,JiangDu,QiJin* StateKeyLaboratoryforMolecularVirologyandGeneticEngineering,InstituteofPathogenBiology,ChineseAcademyofMedicalSciencesandPekingUnionMedical College,Beijing,People’sRepublicofChina Abstract Japanese encephalitis virus (JEV), a neurotropic mosquito-borneflavivirus, causesacute viral encephalitis andneurologic diseasewithahighfatalityrateinhumansandarangeofanimals.SmallinterferingRNA(siRNA)isapowerfulantiviralagent able to inhibit JEV replication. However, the high rate of genetic variability between JEV strains (of four confirmed genotypes,genotypesI,II,IIIandIV)hampersthebroad-spectrumapplicationofsiRNAs,andmutationswithinthetargeted sequences could facilitate JEV escape from RNA interference (RNAi)-mediated antiviral therapy. To improve the broad- spectrumapplicationofsiRNAsandpreventthegenerationofescapemutants,multiplesiRNAstargetingconservedviral sequences need to be combined. In this study, using a siRNA expression vector based on the miR-155 backbone and promotedbyRNApolymeraseII,weinitiallyidentifiedninesiRNAstargetinghighlyconservedregionsofsevenJEVgenes amongstrainsofthefourgenotypesofJEVtoeffectivelyblockthereplicationoftheJEVvaccinestrainSA14-14-2.Then,we constructed single microRNA-like polycistrons to simultaneously express these effective siRNAs under a single RNA polymerase II promoter. Finally, these single siRNAs or multiple siRNAs from the microRNA-like polycistrons showed effectiveanti-virusactivityingenotypeIandgenotypeIIIJEVwildtypestrains,whicharethepredominantgenotypesofJEV in mainlandChina. Theanti-JEV effect ofthese microRNA-like polycistrons was also predictedin othergenotypes ofJEV (genotypes II and IV), The inhibitory efficacy indicated that siRNAs69 could theoretically inhibit the replication of JEV genotypesIIandIV. Citation:WuZ,XueY,WangB,DuJ,JinQ(2011)Broad-Spec