NDLI: Doxycyline regulation in a single retroviral vector by an autoregulatory loop facilitates controlled gene expression in liver cells

Content Provider	PubMed Central
Author	Kühnel, Florian Fritsch, Corinna Krause, Sabine Mundt, Bettina Wirth, Thomas Paul, Yasmin Malek, Nisar Peter Zender, Lars Manns, Michael Peter Kubicka, Stefan
Copyright Year	2004
Abstract	The tetracycline system has limitations in liver cells, such as toxic effects and low controllability. We generated different retroviral vectors for controlled gene expression in liver cells, in which the regulatory elements were arranged in different patterns. Only the organization of the tetracycline system in an autoregulatory loop in the sense orientation results in high retroviral titres and in tight regulation of gene expression in highly differentiated hepatoma cells. Because of the toxicity of the transactivator tTA, it was impossible to establish doxycycline-dependent stable HepG2 cell lines. To avoid sequelching-related toxicity in liver cells, we replaced tTA with new non-toxic transactivators. By using tTA2, tTA3 and tTA4, we observed tight doxycycline-dependent gene expression in 23, 49 and 45% of the isolated clones. The tTA4 vector was used to transduce hepatocytes of mice in vivo. Tight doxycycline-controllable gene regulation was also observed in the liver of mice, confirming our hypothesis that retroviral vectors with autoregulatory loops of the tetracycline system facilitate inducible gene expression in the liver in vivo. Our new retroviral vector system allows rapid isolation of controllable clones in a very high yield and should make the tetracycline system more applicable to liver-derived cells and in liver gene therapy in vivo.
Related Links	http://dx.doi.org/10.1093/nar/gnh034
Ending Page	30
Starting Page	30
File Format	PDF
ISSN	13624962
e-ISSN	13624962
Journal	Nucleic Acids Research
Issue Number	3
Volume Number	32
Language	English
Publisher	Oxford University Press
Publisher Date	2004-02-13
Access Restriction	Open
Rights Holder	Oxford University Press
Subject Keyword	Genetics Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Genetics

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