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在转基因鼠模型中验证猪MyoG启动子驱动猪PPARγ基因的上调表达.docx

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在转基因鼠模型中验证猪MyoG启动子驱动猪PPARγ基因的上调表达 Title:ValidationofporcinePPARγUpregulationDrivenbyPorcineMyoGPromoterinTransgenicMouseModels Abstract: Transgenicanimalmodelshaveproventobevaluabletoolsforunderstandinggeneregulationandstudyinggenefunction.Inthisstudy,weaimedtovalidatetheupregulationofporcinePeroxisomeProliferator-ActivatedReceptorGamma(PPARγ)geneexpressiondrivenbytheporcineMyoGpromoterintransgenicmousemodels.PPARγplaysacriticalroleinadipogenesisandlipidmetabolism,makingitapotentialtargetfortherapeuticinterventionsinobesityandmetabolicdisorders.UnderstandingtheregulationofPPARγexpressioninaporcinemodelwillcontributetothedevelopmentofeffectivestrategiestargetingobesityandrelateddiseases. Introduction: PPARγisamemberofthenuclearreceptorsuperfamilyandactsasakeyregulatorofadipogenesisandlipidmetabolism.Itplaysacrucialroleinthedevelopmentandfunctionofadiposetissue.DysregulationofPPARγexpressionhasbeenimplicatedinvariousmetabolicdisorders,includingobesity,diabetes,andcardiovasculardisease.Therefore,understandingthemechanismsthatcontrolPPARγexpressionisessentialforthedevelopmentoftherapeuticinterventions.TheMyoGpromoter,whichdrivestheexpressionoftheMyoDfamilyoftranscriptionfactors,hasbeenshowntobeactiveduringmyogenesis.However,itspotentialroleinregulatingPPARγexpressionremainstobeexplored. Methods: TovalidatetheupregulationofporcinePPARγexpressiondrivenbytheporcineMyoGpromoter,wegeneratedtransgenicmousemodelscarryingtheporcineMyoGpromoterupstreamoftheporcinePPARγgene.Thetransgenicconstructsweremicroinjectedintofertilizedmouseeggs,andtheresultingfoundermicewereidentifiedbyPCRanalysis.Founderswerethenbredwithwild-typemicetoproduceF1heterozygoustransgenicmice. RNAExtractionandQuantitativeReal-TimePCR(qRT-PCR): TotalRNAwasextractedfromvarioustissuesoftheF1heterozygoustransgenicmiceusingacommercialRNAextractionkit.ReversetranscriptionwasperformedtogeneratecDNA,whichwasthenusedasatemplateforqRT-PCRanalysis.PrimersspecificforporcinePPARγandaninternalreferencegenewereusedtoquantif