Genetic Modification of Cancer Cells Using Non-Viral, Episomal SMAR Vectors for In Vivo Tumour Modelling 英文参考文献.docVIP

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Genetic Modification of Cancer Cells Using Non-Viral, Episomal SMAR Vectors for In Vivo Tumour Modelling 英文参考文献.doc

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Genetic Modification of Cancer Cells Using Non-Viral, Episomal SMAR Vectors for In Vivo Tumour Modelling 英文参考文献

GeneticModificationofCancerCellsUsingNon-Viral, EpisomalS/MARVectorsforInVivoTumourModelling OrestisArgyros.,SuetPingWong.,KateGowers,RichardPaulHarbottle* GeneTherapyResearchGroup,SectionofMolecularMedicine,NationalHeartandLungInstitute,ImperialCollegeLondon,London,UnitedKingdom Abstract Thedevelopmentofgeneticallymarkedanimaltumourxenograftsisanareaofongoingresearchtoenableeasierandmore reliabletestingofcancer therapies.Geneticallymarkedtumourmodels haveanumberofadvantagesoverconventional tumourmodels,includingtheeasylongitudinalmonitoringoftherapiesandthereducednumberofanimalsneededfor trials. Several different methods have been used in previous studies to mark tumours genetically, however all have limitations, such as genotoxicity and other artifacts related to the usage of integrating viral vectors. Recently, we have generatedanepisomallymaintainedplasmidDNA(pDNA)expressionsystembasedonScaffold/MatrixAttachmentRegion (S/MAR),whichpermitslong-termluciferasetransgeneexpressioninthemouseliver.Herewedescribeafurtherusageof thispDNAvectorwiththehumanUbiquitinCpromotertocreatestablytransfectedhumanhepatoma(Huh7)andhuman Pancreatic Carcinoma (MIA-PaCa2) cell lines, which were delivered into ‘‘immune deficient’’ mice and monitored longitudinallyovertimeusingabioluminometer.Bothcelllinesrevealedsustainedepisomallong-termluciferaseexpression and formation of a tumour showing the pathological characteristics of hepatocellular carcinoma (HCC) and pancreatic carcinoma(PaCa),respectively.ThisisthefirstdemonstrationthatapDNAvectorcanconfersustainedepisomalluciferase transgeneexpressioninvariousmousetumourmodelsandcanthusbereadilyutilisedtofollowtumourformationwithout interferingwiththecellulargenome. Citation:ArgyrosO,WongSP,GowersK,HarbottleRP(2012)GeneticModificationofCancerCellsUsingNon-Viral,EpisomalS/MARVectorsforInVivoTumour Modelling.PLoSONE7(10):e47920.doi:10.1371/journal.pone.0047920 Editor:IrinaV.Lebedeva,EnzoLifeSciences,Inc.,UnitedStatesofAmerica ReceivedJun