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A Polyoma-Based Episomal Vector Efficiently Expresses Exogenous Genes in Mouse Embryonic Stem Cells

Camenisch, Gieri ; Gruber, Markus ; Donoho, Greg ; Van Sloun, Petra ; Wenger, Roland H. ; Gassmann, Max

In: Nucleic Acids Research, 1996, vol. 24, no. 19, p. 3707-3713

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    Titel
    • A Polyoma-Based Episomal Vector Efficiently Expresses Exogenous Genes in Mouse Embryonic Stem Cells
    Autor
    • Camenisch, Gieri. Institute of Physiology, University of Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland
    • Gruber, Markus. Institute of Physiology, University of Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland
    • Donoho, Greg. Department of Biochemistry, Beckman Center, Stanford University Medical School, Stanford, CA 94305-5307, USA
    • Van Sloun, Petra. MGC-Department of Radiation Genetics and Chemical Mutagenesis, Leiden University, Wassenaarseweg 72, 2333 AL Leiden, The Netherlands
    • Wenger, Roland H.. Institute of Physiology, University of Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland
    • Gassmann, Max. Institute of Physiology, University of Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland
    Dokumententyp
    • Postprint
    Sprache
    • Englisch
    Veröffentlicht in
    • Nucleic Acids Research, 1996, vol. 24, no. 19, p. 3707-3713. Oxford University Press
    Andere elektronische Ausgabe
    Klassifikation
    • Gesundheit
    OAI-PMH-ID
    • oai:doc.rero.ch:294409
    Summary
    We describe the ability of novel episomally maintained vectors to efficiently promote gene expression in embryonic stem (ES) cells as well as in established mouse cell lines. Extrachromosomal maintenance of our vectors is based on the presence of polyoma virus DNA sequences, including the origin of replication harboring a mutant enhancer (PyF101), and a modified version of the polyoma early region (LT20) encoding the large T antigen only. Reporter gene expression from such extrachromosomally replicating vectors was approximately 10-fold higher than expression from replication-incompetent control plasmids. After transfection of different ES cell lines, the polyoma virus-derived plasmid variant pMGD20neo (7.2 kb) was maintained episomally in 16% of the G418-resistant clones. No chromosomal integration of pMGD20neo vector DNA was detected in ES cells that contained episomal vector DNA even after long term passage. The vector's replication ability was not altered after insertion of up to 10 kb hprt gene fragments. Besides undifferentiated ES cells, the polyoma-based vectors were also maintained extrachromosomally in differentiating ES cells and embryoid bodies as well as in established mouse cell lines