RNAi-Mediated Silencing of Paramyosin Expression in Trichinella spiralis Results in Impaired Viability of the Parasite 英文参考文献.docVIP

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RNAi-Mediated Silencing of Paramyosin Expression in Trichinella spiralis Results in Impaired Viability of the Parasite 英文参考文献.doc

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RNAi-MediatedSilencingofParamyosinExpressionin TrichinellaspiralisResultsinImpairedViabilityofthe Parasite XiaoqinChen,YapingYang,JingYang,ZhifeiZhang,XinpingZhu* DepartmentofParasitology,SchoolofBasicMedicalSciences,CapitalMedicalUniversity,Beijing,China Abstract Background: Trichinella spiralis expresses paramyosin (Ts-PMY) not only as a structural protein but also as an immunomodulatory protein to protect the worm from being attacked by host complement components. In this study, thefunctionsofPMYintheviabilityandthegrowthdevelopmentofT.spiraliswereconfirmedatthefirsttimebysilencing thegenefunctionwithRNAinterferencetechnique. MethodsandFindings:Tounderstanditsfunctionsintheviabilityoftheworm,weusedRNAinterferencetosilencethe expressionofTs-pmymRNAandproteinintheparasite.SignificantsilencingofTs-pmymRNAexpressioninlarvalandadult T.spiraliswasachievedbysiRNAanddsRNAthroughsoakingandelectroporation.ElectroporationofT.spiralislarvaewith 8mMsiRNA1743or100ng/mldsRNA-PF3resultedin66.3%and60.4%decreaseinTs-pmytranscriptand52.0%and64.7% decreaseinTs-PMYproteinexpression,respectively,comparedwithlarvaetreatedwithirrelevantcontrolsiRNAordsRNA. LarvaetreatedwithsiRNA1743displayedsignificantreductioninmolting(40.8%)andserioussurfacedamageasdetected with SYTOX fluorescent staining. Infection of mice with larvae electroporated with Ts-pmy siRNA1743 resulted in 37.6% decrease inadult wormburdenand 23.2%decrease in muscle larvae burdencompared withmice infected withcontrol siRNA-treatedlarvae.Inaddition,adultwormsrecoveredfrommiceinfectedwithsiRNA-treatedlarvaereleased24.8%less newbornlarvae. Conclusion: It is the first time RNAi was used on T. spiralis to demonstrate that silencing PMY expression in T. spiralis significantly reducesthe parasite’s viability andinfectivity,further confirming that Ts-PMY playsanimportant rolein the survivalofT.spiralisandthereforeisapromisingtargetforvaccinedevelopment. Citation:ChenX,YangY,YangJ,ZhangZ,ZhuX(2012)RNAi-MediatedSilencingofParamy