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Titolo:
Generation of a flexible cell line with regulatable, high-level expressionof HIV Gag/Pol particles capable of packaging HIV-derived vectors
Autore:
Sparacio, S; Pfeiffer, T; Schaal, H; Bosch, V;
Indirizzi:
Deutsch Krebsforschungszentrum, Forsch Schwerpunkt Angew Tumorvirol, D-69120 Heidelberg, Germany Deutsch Krebsforschungszentrum Heidelberg Germany D-69120 lberg, Germany Univ Dusseldorf, Inst Med Mikrobiol & Virol, D-4000 Dusseldorf, Germany Univ Dusseldorf Dusseldorf Germany D-4000 ol, D-4000 Dusseldorf, Germany
Titolo Testata:
MOLECULAR THERAPY
fascicolo: 4, volume: 3, anno: 2001,
pagine: 602 - 612
SICI:
1525-0016(200104)3:4<602:GOAFCL>2.0.ZU;2-7
Fonte:
ISI
Lingua:
ENG
Soggetto:
IMMUNODEFICIENCY-VIRUS TYPE-1; VIVO GENE DELIVERY; LENTIVIRUS VECTOR; HISTONE ACETYLATION; PROTEIN EXPRESSION; RETROVIRAL VECTORS; MAMMALIAN-CELLS; SYSTEM; RNA; SEQUENCE;
Keywords:
HIV particles; HIV protease; inducible expression; HIV vector; vector packaging; rev; transduction;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
36
Recensione:
Indirizzi per estratti:
Indirizzo: Bosch, V Deutsch Krebsforschungszentrum, Forsch Schwerpunkt Angew Tumorvirol, Neuenheimer Feld 242,F0200, D-69120 Heidelberg, Germany Deutsch Krebsforschungszentrum Neuenheimer Feld 242,F0200 Heidelberg Germany D-69120
Citazione:
S. Sparacio et al., "Generation of a flexible cell line with regulatable, high-level expressionof HIV Gag/Pol particles capable of packaging HIV-derived vectors", MOL THER, 3(4), 2001, pp. 602-612

Abstract

HIV-derived vectors are of potential clinical relevance due to their ability to transduce nondividing cells in vitro and in vivo. However, the generation of cell lines stably and reproducibly expressing high amounts of defined subviral particles, capable of packaging and transducing HIV-derived vectors, has been hampered by the cytotoxicity of some of the required gene products, in particular of the HIV-1 protease. The successful use of regulatable gene expression systems to overcome this problem requires that the remaining basally expressed gene product activity is below the threshold for cytotoxicity. To try to achieve this, we have consecutively introduced appropriate plasmids, encoding HIV rev and HIV gag/pol gene products, each under the control of separate ecdysone-inducible promoters, into human 293 cells. Using a protocol in which a specific HIV protease inhibitor, Saquinavir, was continuously present in the culture medium during selection, we could generate stable cell lines inducibly expressing high amounts of subviral particles. A cell line, termed 293-Rev/Gag/Pol(i), which has been characterizedin more detail, inducibly releases, within 48 h postinduction, high amounts of HIV Gag/Pol particles (about 10 mug CA/ml). These HIV Gag/Pol particles can package and transduce third-generation HIV vectors to high titers. Thus, in addition to other applications, the 293-Rev/Gag/Pol(i) cell line represents a "founder" packaging cell line which, depending on the requirement, can be further modified to include specific transgene-encoding vector andtargeting glycoprotein genes.

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Documento generato il 09/04/20 alle ore 19:36:11