A 3′-Untranslated Region (3′UTR) Induces Organ Adhesion by Regulating miR-199a Functions 英文参考文献.docVIP

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A 3′-Untranslated Region (3′UTR) Induces Organ Adhesion by Regulating miR-199a Functions 英文参考文献.doc

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A 3′-Untranslated Region (3′UTR) Induces Organ Adhesion by Regulating miR-199a Functions 英文参考文献

A39-UntranslatedRegion(39UTR)InducesOrgan AdhesionbyRegulatingmiR-199a*Functions DanielY.Lee1,2,TatianaShatseva1,2,ZinaJeyapalan1,2,WilliamW.Du1,ZhaoqunDeng1,2,BurtonB. Yang1,2 * 1SunnybrookResearchInstitute,SunnybrookHealthSciencesCentre,Toronto,Canada,2DepartmentofLaboratoryMedicineandPathobiology,UniversityofToronto, Toronto,Canada Abstract Mature microRNAs (miRNAs) are single-stranded RNAs of 18–24 nucleotides that repress post-transcriptional gene expression. However, it is unknown whether the functions of mature miRNAs can be regulated. Here we report that expressionofversican39UTRinducesorganadhesionintransgenicmicebymodulatingmiR-199a*activities.Thestudywas initiatedbythehypothesisthatthenon-coding39UTRplaysaroleintheregulationofmiRNAfunction.Transgenicmice expressingaconstructharboringthe39UTRofversicanexhibitstheadhesionoforgans.Computationalanalysisindicated thatalargenumberofmicroRNAscouldbindtothisfragmentpotentiallyincludingmiR-199a*.Expressionofversicanand fibronectin,twotargetsofmiR-199a*,areup-regulatedintransgenicmice,suggestingthatthe39UTRbindsandmodulates miR-199a*activities, freeing mRNAs ofversicanandfibronectin frombeingrepressed bymiR-199a*. Confirmation ofthe bindingwasperformedbyPCRusingmaturemiR-199a*asaprimerandthetargetingwasperformedbyluciferaseassays. Enhancedadhesionbyexpressionofthe39UTRwasconfirmedbyinvitroassays.Ourresultsdemonstratedthatuponarrival in cytoplasm, miRNA activities can be modulated locally by the 39UTR. Our assay may be developed as sophisticated approachesforstudyingthemutualregulationofmiRNAsandmRNAsinvitroandinvivo.Weanticipatethatexpressionof the39UTRmaybeanapproachinthedevelopmentofgenetherapy. Citation:LeeDY,ShatsevaT,JeyapalanZ,DuWW,DengZ,etal.(2009)A39-UntranslatedRegion(39UTR)InducesOrganAdhesionbyRegulatingmiR-199a* Functions.PLoSONE4(2):e4527.doi:10.1371/journal.pone.0004527 Editor:StefanMaas,LehighUniversity,UnitedStatesofAmerica ReceivedDecember17,2008;AcceptedDecember26,2008;PublishedFebruary18,2009