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Titolo:
Adenoviral vector which delivers FasL-GFP fusion protein regulated by the tet-inducible expression system
Autore:
Rubinchik, S; Ding, R; Qiu, AJ; Zhang, F; Dong, J;
Indirizzi:
Univ Calif San Francisco, Dept Lab Med, San Francisco, CA 94143 USA Univ Calif San Francisco San Francisco CA USA 94143 ancisco, CA 94143 USA Univ Calif San Francisco, Dept Ophthalmol, San Francisco, CA 94143 USA Univ Calif San Francisco San Francisco CA USA 94143 ancisco, CA 94143 USA
Titolo Testata:
GENE THERAPY
fascicolo: 10, volume: 7, anno: 2000,
pagine: 875 - 885
SICI:
0969-7128(200005)7:10<875:AVWDFF>2.0.ZU;2-S
Fonte:
ISI
Lingua:
ENG
Soggetto:
LIGAND GENE-TRANSFER; FACTOR-INDUCED APOPTOSIS; IMMUNE PRIVILEGE; NECROSIS-FACTOR; RECOMBINANT ADENOVIRUS; MEDIATED CYTOTOXICITY; TRANSGENE EXPRESSION; ALLOGRAFT SURVIVAL; MAMMALIAN-CELLS; TUMOR-CELLS;
Keywords:
Fas-ligand; apoptosis; adenoviral vector; regulation; regulated expression;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
47
Recensione:
Indirizzi per estratti:
Indirizzo: Dong, J Med Univ S Carolina, Dept Microbiol & Immunol, BSB201,173 Ashley Rd, Charleston, SC 29425 USA Med Univ S Carolina BSB201,173 Ashley Rd Charleston SC USA 29425 A
Citazione:
S. Rubinchik et al., "Adenoviral vector which delivers FasL-GFP fusion protein regulated by the tet-inducible expression system", GENE THER, 7(10), 2000, pp. 875-885

Abstract

Fas ligand (FasL) is a member of the tumor necrosis family and when bound to its receptor, Fas, induces apoptosis. It plays important roles in immuneresponse, degenerative and lymphoproliferative diseases, development and tumorigenesis. It is also involved in generation of immune privilege sites in the eye and testis. Harnessing the power of this molecule is expected to lead to a powerful chemotherapeutic. We describe the construction and characterization of replication-deficient adenoviral vectors that express a fusion of murine FasL and green fluorescent protein (GFP). FasL-GFP retains full activity of wild-type FasL, at the same time allowing for easy visualization and quantification in both living and fixed cells. The fusion protein is under the control of a tetracycline-regulated gene expression system. Tight control of expression is achieved by creating a novel 'double recombinant' Ad vector in which the tet-responsive element and the transactivator element are built into the opposite ends of the same vector to avoid enhancer interference. Expression can be conveniently regulated by tetracycline or its derivatives in a dose-dependent manner. The vector was able to deliver FasL-GFP gene to cells in vitro efficiently, and the expression level and function of the fusion protein was modulated by the concentration of doxycycline. This regulation allows us to produce high titers of the vector by inhibiting FasL expression in an apoptosis-resistant cell line. Induction of apoptosis was demonstrated in all cell lines tested. These results indicate that our vector is a potentially valuable tool for FasL-based gene therapy of cancer and for the study of FasL/Fas-mediated apoptosis and immune privilege.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 01/04/20 alle ore 23:46:41