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Titolo:
cis requirements for the efficient production of recombinant DNA vectors based on autonomous parvoviruses
Autore:
Kestler, J; Neeb, B; Struyf, S; Van Damme, J; Cotmore, SF; DAbramo, A; Tattersall, P; Rommelaere, J; Dinsart, C; Cornelis, JJ;
Indirizzi:
Deutsch Krebsforschungszentrum, Appl Tumor Virol Abt F0100, D-69009 Heidelberg, Germany Deutsch Krebsforschungszentrum Heidelberg Germany D-69009 lberg, Germany Deutsch Krebsforschungszentrum, INSERM, U375, D-69009 Heidelberg, Germany Deutsch Krebsforschungszentrum Heidelberg Germany D-69009 lberg, Germany Catholic Univ Louvain, Rega Inst, Lab Mol Immunol, B-3000 Louvain, BelgiumCatholic Univ Louvain Louvain Belgium B-3000 ol, B-3000 Louvain, Belgium Yale Univ, Sch Med, Dept Lab Med, New Haven, CT 06510 USA Yale Univ New Haven CT USA 06510 d, Dept Lab Med, New Haven, CT 06510 USA Yale Univ, Sch Med, Dept Genet, New Haven, CT 06510 USA Yale Univ New Haven CT USA 06510 Med, Dept Genet, New Haven, CT 06510 USA
Titolo Testata:
HUMAN GENE THERAPY
fascicolo: 10, volume: 10, anno: 1999,
pagine: 1619 - 1632
SICI:
1043-0342(19990701)10:10<1619:CRFTEP>2.0.ZU;2-5
Fonte:
ISI
Lingua:
ENG
Soggetto:
TRANSFORMED RAT-CELLS; MINUTE VIRUS; NONSTRUCTURAL PROTEINS; GENE-THERAPY; ADENOASSOCIATED VIRUS; MICE MINIGENOMES; EPITHELIAL-CELLS; NS1 POLYPEPTIDE; P38 PROMOTER; JE GENE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
49
Recensione:
Indirizzi per estratti:
Indirizzo: Cornelis, JJ Deutsch Krebsforschungszentrum, Appl Tumor Virol Abt 0610, Postfach 101949, D-69009 Heidelberg, Germany Deutsch Krebsforschungszentrum Postfach 101949 Heidelberg Germany D-69009
Citazione:
J. Kestler et al., "cis requirements for the efficient production of recombinant DNA vectors based on autonomous parvoviruses", HUM GENE TH, 10(10), 1999, pp. 1619-1632

Abstract

The replication of viral genomes and the production of recombinant viral vectors from infectious molecular clones of parvoviruses MVMp and H1 were greatly improved by the introduction of a consensus NS-1 nick site at the junction between the left-hand viral terminus and the plasmid DNA, Progressivedeletions of up to 1600 bp in the region encoding the structural genes as well as insertions of foreign DNA in replacement of those sequences did notappreciably affect the replication ability of the recombinant H1 virus genomes, In contrast, the incorporation of these genomes into recombinant particles appeared to depend on in cis-provided structural gene sequences. Indeed, the production of H1 viral vectors by cotransfection of recombinant clones and helper plasmids providing the structural proteins (VPs) in trans, drastically decreased when more than 800 bp was removed from the VP transcription unit. Furthermore, titers of viral vectors, in which most of the VP-coding region was replaced by an equivalent-length sequence consisting of reporter cDNA and stuffer DNA, were reduced more than 50 times in comparison with recombinant vectors in which stuffer DNA was not substituted for the residual VP sequence. In addition, viral vector production was restricted bythe overall size of the genome, with a mere 6% increase in DNA length leading to an approximately 10 times lower encapsidation yield. Under conditions fulfilling the above-mentioned requirements for efficient packaging, titers of virus vectors from improved recombinant molecular DNA clones amountedto 5 x 10(7) infectious units per milliliter of crude extract. These titers should allow the assessment of the therapeutic effect of recombinant parvoviruses expressing small transgenes in laboratory animals.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 04/12/20 alle ore 01:30:46