title: Tet system in the brain: transgenic rats and lentiviral vectors approach.
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.
authors.affiliation: Department of Molecular and Cellular Neurobiology, Nencki Institute, Pasteura 3, Warsaw, Poland. w.konopka@dkfz.de
authors.forename: Witold
authors.initials: W
authors.lastname: Konopka
authors.forename: Kamila
authors.initials: K
authors.lastname: Duniec
authors.forename: Agata
authors.initials: A
authors.lastname: Klejman
authors.forename: Marcin
authors.initials: M
authors.lastname: Wawrzyniak
authors.forename: Dorota
authors.initials: D
authors.lastname: Owczarek
authors.forename: Ludwika
authors.initials: L
authors.lastname: Gawrys
authors.forename: Marek
authors.initials: M
authors.lastname: Maleszewski
authors.forename: Jacques
authors.initials: J
authors.lastname: Mallet
authors.forename: Leszek
authors.initials: L
authors.lastname: Kaczmarek
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000378:metabolism*; D002478:Cells, Cultured; D018891:Dentate Gyrus / Q000378:metabolism; D004318:Doxycycline / Q000494:pharmacology; D005260:Female; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D016086:Lentivirus / Q000235:genetics; D008297:Male; D051381:Rats; D055647:Rats, Transgenic; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020218:Response Elements / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013752:Tetracycline / Q000494:pharmacology; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D049452:Green Fluorescent Proteins; D013752:Tetracycline; D004318:Doxycycline