RNAi Methodologies for the Functional Study of Signaling Molecules 英文参考文献.docVIP

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RNAi Methodologies for the Functional Study of Signaling Molecules 英文参考文献.doc

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RNAi Methodologies for the Functional Study of Signaling Molecules 英文参考文献

RNAiMethodologiesfortheFunctionalStudyof SignalingMolecules GwangLee1,2,LeahA.Santat3,MiSookChang3,SangdunChoi1,4* 1DepartmentofMolecularScienceandTechnology,CollegeofNaturalScience,AjouUniversity,Suwon,Korea,2InstituteforMedicalSciences,CollegeofNaturalScience, AjouUniversity,Suwon,Korea,3MolecularBiologyLaboratory,AllianceforCellularSignaling,DivisionofBiology,CaliforniaInstituteofTechnology,Pasadena,California, UnitedStatesofAmerica,4DepartmentofBiologicalSciences,CollegeofNaturalScience,AjouUniversity,Suwon,Korea Abstract RNAinterference(RNAi)wasinvestigatedwiththeaimofachievinggenesilencingwithdiverseRNAiplatformsthatinclude smallinterferingRNA(siRNA),shorthairpinRNA(shRNA)andantisenseoligonucleotides(ASO).Differentversionsofeach systemwereusedtosilencetheexpressionofspecificsubunitsoftheheterotrimericsignaltransducingG-proteins,Galpha i2 and G beta 2, in the RAW 264.7 murine macrophage cell line. The specificity of the different RNA interference (RNAi) platformswasassessedbyDNAmicroarrayanalysis.ReliableRNAimethodologiesagainstthegenesofinterestwerethen developed and applied to functional studies of signaling networks. This study demonstrates a successful knockdown of targetgenesandshowsthepotentialofRNAiforuseinfunctionalstudiesofsignalingmolecules. Citation:LeeG,SantatLA,ChangMS,ChoiS(2009)RNAiMethodologiesfortheFunctionalStudyofSignalingMolecules.PLoSONE4(2):e4559.doi:10.1371/ journal.pone.0004559 Editor:TerryMeans,MassachusettsGeneralHospital/HarvardUniversity,UnitedStatesofAmerica ReceivedJanuary2,2009;AcceptedJanuary13,2009;PublishedFebruary24,2009 Copyright:?2009Leeetal.Thisisanopen-accessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,whichpermitsunrestricted use,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:ThisworkwassupportedbyaKoreaScienceandEngineeringFoundation(KOSEF)grantfundedbytheKoreanGovernment(MOST)(R01-2007-000- 20533-0).ThisworkwasalsopartlysupportedbyaKoreaResearchF