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dc.contributor.authorGama-Norton, Leonor
dc.contributor.authorBotezatu, Lacramioara
dc.contributor.authorHerrmann, Sabrina
dc.contributor.authorSchweizer, Matthias
dc.contributor.authorAlves, Paula Marques
dc.contributor.authorHauser, Hansjoerg
dc.contributor.authorWirth, Dagmar
dc.date.accessioned2012-08-02T09:04:32Z
dc.date.available2012-08-02T09:04:32Z
dc.date.issued2011-10
dc.identifier.citationLentivirus production is influenced by SV40 large T-antigen and chromosomal integration of the vector in HEK293 cells. 2011, 22 (10):1269-79 Hum. Gene Ther.en_GB
dc.identifier.issn1557-7422
dc.identifier.pmid21554103
dc.identifier.doi10.1089/hum.2010.143
dc.identifier.urihttp://hdl.handle.net/10033/236995
dc.description.abstractCurrently, lentiviral vectors for research and gene therapy are produced from 293-T cells that are transiently transfected with plasmids encoding the vector and helper functions. However, transiently transfected vectors as well as the presence of SV40 virus large T-antigen (T-Ag) cause serious technical and safety considerations. We aimed to exploit single copy integration sites in the HEK293 genome supporting lentiviral vector production. We found that lentiviral vectors result in minimal infectious particle production from single copy integrants in HEK293. Moreover, once this cell line harbors single copy integrations of lentiviral vectors, its ability to transiently produce lentiviral vectors becomes strongly impaired. T-Ag has a dramatic effect on virus production. Low levels of constitutive T-Ag expression can overcome the production restriction imposed by integrated lentiviral vectors copies. Interestingly, T-Ag does not exert its role at the level of transcriptional activity of the vector; rather, it seems to impose an indirect effect on the cell thereby enabling lentiviral vector production. Altogether, our study highlights the restrictions for integrated lentiviral vectors that are relevant for the establishment of stable and safe producer cell lines.
dc.language.isoenen
dc.rightsArchived with thanks to Human gene therapyen_GB
dc.subject.meshAntigens, Polyomavirus Transformingen_GB
dc.subject.meshChromosomes, Humanen_GB
dc.subject.meshDNA Primersen_GB
dc.subject.meshFlow Cytometryen_GB
dc.subject.meshGenetic Vectorsen_GB
dc.subject.meshHEK293 Cellsen_GB
dc.subject.meshHumansen_GB
dc.subject.meshLentivirusen_GB
dc.subject.meshReal-Time Polymerase Chain Reactionen_GB
dc.subject.meshTransduction, Geneticen_GB
dc.subject.meshTransfectionen_GB
dc.subject.meshVirus Integrationen_GB
dc.titleLentivirus production is influenced by SV40 large T-antigen and chromosomal integration of the vector in HEK293 cells.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Centre for Infection Research, Braunschweig, Germany .en_GB
dc.identifier.journalHuman gene therapyen_GB
refterms.dateFOA2018-06-12T18:10:24Z
html.description.abstractCurrently, lentiviral vectors for research and gene therapy are produced from 293-T cells that are transiently transfected with plasmids encoding the vector and helper functions. However, transiently transfected vectors as well as the presence of SV40 virus large T-antigen (T-Ag) cause serious technical and safety considerations. We aimed to exploit single copy integration sites in the HEK293 genome supporting lentiviral vector production. We found that lentiviral vectors result in minimal infectious particle production from single copy integrants in HEK293. Moreover, once this cell line harbors single copy integrations of lentiviral vectors, its ability to transiently produce lentiviral vectors becomes strongly impaired. T-Ag has a dramatic effect on virus production. Low levels of constitutive T-Ag expression can overcome the production restriction imposed by integrated lentiviral vectors copies. Interestingly, T-Ag does not exert its role at the level of transcriptional activity of the vector; rather, it seems to impose an indirect effect on the cell thereby enabling lentiviral vector production. Altogether, our study highlights the restrictions for integrated lentiviral vectors that are relevant for the establishment of stable and safe producer cell lines.


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