RNA Modulators of Complex Phenotypes in Mammalian Cells 英文参考文献.docVIP

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RNA Modulators of Complex Phenotypes in Mammalian Cells 英文参考文献.doc

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RNA Modulators of Complex Phenotypes in Mammalian Cells 英文参考文献

RNAModulatorsofComplexPhenotypesinMammalian Cells AngelaLai1¤a,MurrayJ.Cairns1¤b,NhamTran1,2¤c,Hong-PingZhang1,LaraCullen1¤a,GregM.Arndt1* 1JohnsonandJohnsonResearchPtyLtd,AustralianTechnologyPark,Eveleigh,NewSouthWales,Australia,2TheSydneyHeadandNeckCancerInstitute,SydneyCancer Centre,RoyalPrinceAlfredHospitalandUniversityofSydney,Sydney,NewSouthWales,Australia Abstract RNA-mediatedgenesilencing,intheformofRNAinterference(RNAi)ormicroRNAs(miRNAs)hasprovidednoveltoolsfor genediscoveryandvalidationinmammaliancells.Here,wereportontheconstructionandapplicationofarandomsmall RNA expression library for use in identifying small interfering RNA (siRNA) effectors that can modify complex cellular phenotypes in mammalian cells. The library is based in a retroviral vector and uses convergent promoters to produce unique small complementary RNAs. Using this library, we identify a range of small RNA-encoding gene inserts that overcomeresistanceto5-fluorouracil(5-FU)-ortumournecrosisfactoralpha(TNF-a)-inducedcelldeathincolorectalcancer cells.WedemonstratetheutilityofthistechnologyplatformbyidentifyingakeyRNAeffector,intheformofasiRNA,which overcomescelldeathinducedbythechemotherapeutic5-FU.Thetechnologydescribedhasthepotentialtoidentifyboth functional RNA modulators capable of altering physiological systems and the cellular target genes altered by these modulators. Citation: Lai A, Cairns MJ, Tran N, Zhang H-P, Cullen L, et al. (2009) RNA Modulators of Complex Phenotypes in Mammalian Cells. PLoS ONE 4(3): e4758. doi:10.1371/journal.pone.0004758 Editor:ThomasPreiss,VictorChangCardiacResearchInstitute(VCCRI),Australia ReceivedDecember11,2008;AcceptedFebruary12,2009;PublishedMarch9,2009 Copyright:?2009Laietal.Thisisanopen-accessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,whichpermitsunrestricted use,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:FinancialsupportforthisworkwasprovidedbyJohnsonandJohnsonResearc