Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
Efficient production of adeno-associated virus vectors using split-type helper plasmids
Autore:
Ogasawara, Y; Urabe, M; Kogure, K; Kume, A; Colosi, P; Kurtzman, GJ; Ozawa, K;
Indirizzi:
Jichi90498,ch, Ctr Mol Med, Div Genet Therapeut, Minami Kawachi, Tochigi 32 Jichi Med Sch Minami Kawachi Tochigi Japan 3290498 mi Kawachi, Tochigi 32 Japan Sci & Technol Corp, CREST, Kawaguchi, Saitama 3320012, Japan Japan Sci & Technol Corp Kawaguchi Saitama Japan 3320012 a 3320012, Japan Jikei Univ, Sch Med, Dept Internal Med 2, Minato Ku, Tokyo 1050003, Japan Jikei Univ Tokyo Japan 1050003 al Med 2, Minato Ku, Tokyo 1050003, Japan Avigen Inc, Alameda, CA 94502 USA Avigen Inc Alameda CA USA 94502Avigen Inc, Alameda, CA 94502 USA
Titolo Testata:
JAPANESE JOURNAL OF CANCER RESEARCH
fascicolo: 4, volume: 90, anno: 1999,
pagine: 476 - 483
SICI:
0910-5050(199904)90:4<476:EPOAVV>2.0.ZU;2-9
Fonte:
ISI
Lingua:
ENG
Soggetto:
RECOMBINANT ADENOASSOCIATED VIRUS; REP GENE; CELL-LINES; REPLICATION; EXPRESSION; DNA; PROTEINS; THERAPY; MUTANTS; AAV;
Keywords:
AAV vector; helper plasmid; Rep; Cap; gene therapy;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
32
Recensione:
Indirizzi per estratti:
Indirizzo: Ozawa, K Jichiawachi,h, Ctr Mol Med, Div Genet Therapeut, 3311-1 Yakushiji, Minami K Jichi Med Sch 3311-1 Yakushiji Minami Kawachi Tochigi Japan 3290498
Citazione:
Y. Ogasawara et al., "Efficient production of adeno-associated virus vectors using split-type helper plasmids", JPN J CANC, 90(4), 1999, pp. 476-483

Abstract

Adeno-associated virus (AAV) vectors are potentially useful vehicles for the delivery of therapeutic genes into human cells. To determine the optimalexpression pattern of AAV proteins (Rep78, Rep68, Rep52, Rep40, and Cap proteins) for packaging the recombinant AAV genome, helper plasmids were split into two portions. In this study, two sets of split-type helper plasmids were prepared: i.e., 1) a Rep expression plasmid (pR78/68) and Cap expression plasmid (pCap), and 2) a large Rep expression plasmid (pR78/68 and smallRep plus Cap expression plasmid (pR52/40Cap). When AAV vectors were produced using these sets of split-type helper plasmids at various ratios, the optimal ratio of (large) Rep expression plasmid and Cap expression plasmid was 1 to 9 for both sets. More importantly, the titers were comparable to or even higher than that of a conventional helper plasmid (pIM45) (4.9+/-2.1x10(11) vector particles/10 cm dish for pRep and pCap; 2.9+/-1.6x10(11) vector particles/10 cm dish for pR78/68 and pR52/40Cap: and 1.8+/-0.16x10(11) particles/10 cm dish for pIM45). Western analysis of AAV proteins suggests that the expression of a relatively small amount of large rep and a large amount of Cap is important for optimal vector production. The present study shows that the AAV helper plasmid can be split without losing the ability to package the recombinant AAV genome, and provides us with valuable basic information for the development of efficient AAV packaging cell lines.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 04/06/20 alle ore 20:41:40