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Tet system in the brain: Transgenic rats and lentiviral vectors approach
Author(s) -
Konopka Witold,
Duniec Kamila,
Klejman Agata,
Wawrzyniak Marcin,
Owczarek Dorota,
Gawrys Ludwika,
Maleszewski Marek,
Mallet Jacques,
Kaczmarek Leszek
Publication year - 2009
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.20487
Subject(s) - transgene , biology , doxycycline , dentate gyrus , viral vector , transgenesis , reporter gene , green fluorescent protein , genetically modified mouse , microbiology and biotechnology , gene , gene expression , activator (genetics) , central nervous system , neuroscience , genetics , recombinant dna , antibiotics , reproductive biology , embryogenesis
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. genesis 47:274–280, 2009. © 2009 Wiley‐Liss, Inc.