Tetracycline-inducible transgene expression mediated by a single AAV vector.

Détails

ID Serval
serval:BIB_19117154A560
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Tetracycline-inducible transgene expression mediated by a single AAV vector.
Périodique
Gene Therapy
Auteur(s)
Chtarto A., Bender H.U., Hanemann C.O., Kemp T., Lehtonen E., Levivier M., Brotchi J., Velu T., Tenenbaum L.
ISSN
0969-7128 (Print)
ISSN-L
0969-7128
Statut éditorial
Publié
Date de publication
2003
Peer-reviewed
Oui
Volume
10
Numéro
1
Pages
84-94
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Résumé
Regulated gene delivery systems are usually made of two elements: an inducible promoter and a transactivator. In order to optimize gene delivery and regulation, a single viral vector ensuring adequate stoichiometry of the two elements is required. However, efficient regulation is hampered by interferences between the inducible promoter and (i) the promoter used to express the transactivator and/or (ii) promoter/enhancer elements present in the viral vector backbone. We describe a single AAV vector in which transcription of both the reverse tetracycline transactivator (rtTA) and the transgene is initiated from a bidirectional tetracycline-responsive promoter and terminated at bidirectional SV40 polyadenylation sites flanking both ITRs. Up to 50-fold induction of gene expression in human tumor cell lines and 100-fold in primary cultures of rat Schwann cells was demonstrated. In addition an 80-fold induction in vivo in the rat brain has been obtained. In vitro, the autoregulatory vector exhibits an induced expression level superior to that obtained using the constitutive CMV promoter. Although extinction of the transgene after removal of tetracycline was rapid (less than 3 days), inducibility after addition of tetracycline was slow (about 14 days). This kinetics is suitable for therapeutic gene expression in slowly progressive diseases while allowing rapid switch-off in case of undesirable effects. As compared to previously described autoregulatory tet-repressible (tetOFF) AAV vectors, the tet-inducible (tetON) vector prevents chronic antibiotic administration in the uninduced state.
Mots-clé
Animals, Anti-Bacterial Agents/metabolism, Anti-Bacterial Agents/therapeutic use, Cells, Cultured, Dependovirus/genetics, Entopeduncular Nucleus/metabolism, Flow Cytometry, Gene Expression Regulation, Genetic Engineering, Genetic Therapy/methods, Genetic Vectors/administration & dosage, Genetic Vectors/genetics, Green Fluorescent Proteins, HeLa Cells, Humans, Luminescent Proteins/genetics, Microscopy, Fluorescence, Rats, Schwann Cells/metabolism, Tetracycline/metabolism, Tetracycline/therapeutic use, Transfection/methods, Transgenes, Tumor Cells, Cultured, Virus Diseases/therapy
Pubmed
Web of science
Open Access
Oui
Création de la notice
20/01/2008 17:35
Dernière modification de la notice
20/08/2019 12:49
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