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Nucleic Acids Research 2004 32(18):5529-5538; doi:10.1093/nar/gkh887
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Published online 14 October 2004

Nucleic Acids Research, Vol. 32 No. 18 © Oxford University Press 2004; all rights reserved

Transcriptional control of SV40 T-antigen expression allows a complete reversion of immortalization

Tobias May, Hansjörg Hauser* and Dagmar Wirth

Department of Gene Regulation and Differentiation, GBF—German Research Center for Biotechnology, Mascheroder Weg 1, D-38124 Braunschweig, Germany

* To whom correspondence should be addressed. Tel: +49 531 6181 250; Fax: +49 531 6181 262; Email: hha{at}gbf.de

Received July 8, 2004; Revised and Accepted September 24, 2004

Conditional proliferation of mouse embryo fibroblasts was achieved with a novel autoregulatory vector for Tet-dependent expression of the SV40 T-antigen. The majority of cell clones that were isolated under induced conditions showed strict regulation of cell growth. Status switches were found to be fully reversible and highly reproducible with respect to gene expression characteristics. A consequence of T-antigen expression is a significant deregulation of >400 genes. Deinduced cells turn to rest in G0/G1 phase and exhibit a senescent phenotype. The cells are not oncogenic and no evidence for transformation was found after several months of cultivation. Conditional immortalization allows diverse studies including those on cellular activities without the influence of the immortalizing gene(s), senescence as well as secondary effects from T-antigen expression.


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