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. 2009 Apr;47(4):274-80.
doi: 10.1002/dvg.20487.

Tet system in the brain: transgenic rats and lentiviral vectors approach

Witold Konopka  1 , Kamila Duniec, Agata Klejman, Marcin Wawrzyniak, Dorota Owczarek, Ludwika Gawrys, Marek Maleszewski, Jacques Mallet, Leszek Kaczmarek
Affiliations

Tet system in the brain: transgenic rats and lentiviral vectors approach

Witold Konopka et al. Genesis. 2009 Apr.

Abstract

Local and regulated expression of exogenous genes in the central nervous system is one of the major challenges of modern neuroscience. We have approached this issue by applying the inducible tetracycline system to regulate the expression of EGFP reporter gene in double transgenic rats. We have obtained a strong induction of EGFP only in male testes, which correlated with a high level of rtTA expression only in this organ. To overcome the problem of lack of rtTA protein in the transgenic rat brain, we have delivered this Tet system activator with lentiviral vectors into the dentate gyrus of hippocampus of transgenic EGFP rats. As a result, after systemic application of doxycycline we have obtained inducible, stable and restricted to the desired brain region expression of EGFP. An advantage of this strategy is that the transgene is located in the same genetic milieu in every cell of the transgenic organism. This is crucial to obtain uniform expression of the regulated gene within the target brain structure. Combination of rat transgenesis and lentiviral vectors is a novel approach enabling precise spatiotemporal regulation of genes of interest strictly in the brain structure of choice or in other tissues.

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