Rat tyrosine hydroxylase promoter directs tetracycline-inducible foreign gene expression in dopaminergic cell types

Brain Research. Molecular Brain Research
Mossa Gardaneh, K L O'Malley

Abstract

A prerequisite for creating animal models in which gene expression is spatially and temporally controlled is the development of promoters to target genetic switches to specific populations of cells. Here we used the dopaminergic biosynthetic enzyme, tyrosine hydroxylase (TH) to test various combinations of tetracycline (Tet) system elements to determine the optimal configuration for inducible, tissue-specific expression. The present study shows that the degree of expression and level of leakiness associated with the Tet transactivators rtTA, rtTA2S-M2, tTS/rtTA or tTS/rtTA2S-M2 was dependent upon both the promoter and cell type utilized. Specifically, CMV-driven tTS/rtTA2S-M2 exhibited the highest level of inducibility in HEK cells (approximately 1000-fold) versus the dopaminergic cell line, MN9D (approximately 70-fold). In contrast, TH-driven rtTA2S-M2 yielded the highest level of expression with the least background in dopaminergic cell types versus HEK cells. Moreover, the TH promoter could be combined with the bi-directional Tet response system, BiTetO, allowing for the co-expression and regulation of two genes in the same cell. To further test the feasibility of this system we replaced the reporter gene with human Bcl-2. C...Continue Reading

References

Jun 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·M Gossen, H Bujard
Jun 22, 2000·Proceedings of the National Academy of Sciences of the United States of America·S UrlingerW Hillen
Oct 5, 2001·Nature Reviews. Genetics·M Lewandoski
Jan 29, 2002·Human Gene Therapy·Stefania LamartinaCarlo Toniatti
Jul 20, 2002·Seminars in Cell & Developmental Biology·Zhou ZhuJack A Elias
Sep 19, 2003·Neurobiology of Disease·Karen L O'MalleyWilliam Holtz

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Citations

Dec 22, 2006·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Krish ChandrasekaranTibor Kristian
Nov 9, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Isabelle BardeOlivier Danos

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