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Retroviral vectors for establishing tetracycline-regulated gene expression in an otherwise recalcitrant cell line

Kenny, PA; Enver, T; Ashworth, A; (2002) Retroviral vectors for establishing tetracycline-regulated gene expression in an otherwise recalcitrant cell line. BMC MOL BIOL , 3 , Article 13. 10.1186/1471-2199-3-13. Green open access

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Abstract

Background: Tetracycline-regulated systems have been used to control the expression of heterologous genes in such diverse organisms as yeast, plants, flies and mice. Adaptation of this prokaryotic regulatory system avoids many of the problems inherent in other inducible systems. There have, however, been many reports of difficulties in establishing functioning stable cell lines due to the cytotoxic effects of expressing high levels of the tetracycline transactivator, tTA, from a strong viral promoter.Results: Here we report the successful incorporation of tetracycline-mediated gene expression in a mouse mammary epithelial cell line, HC11, in which conventional approaches failed. We generated retroviruses in which tTA expression was controlled by one of three promoters: a synthetic tetracycline responsive promoter (TRE), the elongation factor 1-alpha promoter (EF1alpha) or the phosphoglycerate kinase-1 promoter (PGK), and compared the resulting cell lines to one generated using a cytomegalovirus immediate early gene promoter (CMV). In contrast to cells produced using the CMV and PGK promoters, those produced using the EF1alpha and TRE promoters expressed high levels of beta-galactosidase in a tetracycline-dependent manner.Conclusions: These novel retroviral vectors performed better than the commercially available system and may have a more general utility in similarly recalcitrant cell lines.

Type: Article
Title: Retroviral vectors for establishing tetracycline-regulated gene expression in an otherwise recalcitrant cell line
Open access status: An open access version is available from UCL Discovery
DOI: 10.1186/1471-2199-3-13
Publisher version: http://www.biomedcentral.com/1471-2199/3/13
Language: English
Additional information: © 2002 Kenny et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
Keywords: MAMMALIAN-CELLS, TRANSGENIC MICE, TRANSCRIPTIONAL ACTIVATOR, SYSTEM, PROMOTER, REPRESSOR, TRANSFECTION, RESISTANCE, PROTEINS, GAL4
UCL classification: UCL
UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute > Research Department of Cancer Bio
URI: https://discovery.ucl.ac.uk/id/eprint/1336580
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