2.50
Hdl Handle:
http://hdl.handle.net/10033/236995
Title:
Lentivirus production is influenced by SV40 large T-antigen and chromosomal integration of the vector in HEK293 cells.
Authors:
Gama-Norton, Leonor; Botezatu, Lacramioara; Herrmann, Sabrina; Schweizer, Matthias; Alves, Paula Marques; Hauser, Hansjoerg; Wirth, Dagmar
Abstract:
Currently, 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.
Affiliation:
Helmholtz Centre for Infection Research, Braunschweig, Germany .
Citation:
Lentivirus 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.
Journal:
Human gene therapy
Issue Date:
Oct-2011
URI:
http://hdl.handle.net/10033/236995
DOI:
10.1089/hum.2010.143
PubMed ID:
21554103
Type:
Article
Language:
en
ISSN:
1557-7422
Appears in Collections:
Publications of AG Modellsysteme für Infektion und Immunität (MSYS)

Full metadata record

DC FieldValue Language
dc.contributor.authorGama-Norton, Leonoren_GB
dc.contributor.authorBotezatu, Lacramioaraen_GB
dc.contributor.authorHerrmann, Sabrinaen_GB
dc.contributor.authorSchweizer, Matthiasen_GB
dc.contributor.authorAlves, Paula Marquesen_GB
dc.contributor.authorHauser, Hansjoergen_GB
dc.contributor.authorWirth, Dagmaren_GB
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.en_GB
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

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