Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
Improved helper virus-free packaging system for HSV amplicon vectors usingan ICP27-deleted, oversized HSV-1 DNA in a bacterial artificial chromosome
Autore:
Saeki, Y; Fraefel, C; Ichikawa, T; Breakefield, XO; Chiocca, EA;
Indirizzi:
Harvard Univ, Mol Neurooncol Labs, Massachusetts Gen Hosp, Sch Med,Neurosurg Serv, Charlestown, MA 02129 USA Harvard Univ Charlestown MA USA 02129 urg Serv, Charlestown, MA 02129 USA Harvard Univ, Dept Neurol, Massachusetts Gen Hosp, Sch Med,Mol Neurogenet Unit, Charlestown, MA 02129 USA Harvard Univ Charlestown MA USA 02129 net Unit, Charlestown, MA 02129 USA Osaka Univ, Grad Sch Med, Div Gene Therapy Sci, Suita, Osaka 5650871, Japan Osaka Univ Suita Osaka Japan 5650871 apy Sci, Suita, Osaka 5650871, Japan Univ Zurich, Inst Virol, CH-8057 Zurich, Switzerland Univ Zurich Zurich Switzerland CH-8057 irol, CH-8057 Zurich, Switzerland
Titolo Testata:
MOLECULAR THERAPY
fascicolo: 4, volume: 3, anno: 2001,
pagine: 591 - 601
SICI:
1525-0016(200104)3:4<591:IHVPSF>2.0.ZU;2-Q
Fonte:
ISI
Lingua:
ENG
Soggetto:
EPSTEIN-BARR-VIRUS; ESCHERICHIA-COLI; TETRACYCLINE RESISTANCE; GENE-TRANSFER; NEURAL CELLS; IN-VIVO; SIMPLEX; TYPE-1; EXPRESSION; REPLICATION;
Keywords:
herpes virus; amplicon; helper virus-free; BAC; homologous recombination; site-specific recombination; gene therapy; RecA; Cre/loxP;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
30
Recensione:
Indirizzi per estratti:
Indirizzo: Saeki, Y Harvard Univ, Mol Neurooncol Labs, Massachusetts Gen Hosp, Sch Med,Neurosurg Serv, CNY6119,149 13th St, Charlestown, MA 02129 USA Harvard Univ CNY6119,149 13th St Charlestown MA USA 02129 129 USA
Citazione:
Y. Saeki et al., "Improved helper virus-free packaging system for HSV amplicon vectors usingan ICP27-deleted, oversized HSV-1 DNA in a bacterial artificial chromosome", MOL THER, 3(4), 2001, pp. 591-601

Abstract

Herpes simplex virus type 1 (HSV-1) amplicons are prokaryotic plasmids containing one or more transcriptional units and two cis-acting HSV-1 sequences: a viral origin of DNA replication and a viral DNA cleavage/packaging signal. In the presence of HSV-1 "helper" functions, amplicons are replicated and packaged into HSV-1 virions. Despite recent improvements in packaging methods, stocks of amplicon vectors are still contaminated with replication-competent helper virus at a frequency of 10(-4)-10(-6). TO overcome this problem, we report that: (i) genetic modifications of HSV-1 genomes can be routinely achieved in Escherichia coli, either by homologous or site-specificrecombination, (ii) a novel HSV-1 bacterial artificial chromosome (fHSV Delta pac27 0+), which has a deletion in the essential gene encoding ICP27 and an addition of ICP0 "stuffer" sequences to increase its size to 178 kb, supports the replication and packaging of cotransfected amplicon DNA withoutgenerating replication-competent helper virus (<1 helper virus per 10(8) TU amplicon vectors), and (iii) the resulting amplicon stocks have titers ofup to 3-10 x 10(8) TU/ml after concentration. Elimination of replication-competent helper virus from HSV-1 amplicon vector stocks further improves safety in gene transfer applications.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 26/02/20 alle ore 05:18:01