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Transposon-assisted cloning and traceless mutagenesis of adenoviruses: Development of a novel vector based on species D

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Ruzsics, Zsolt, Wagner, Markus, Osterlehner, Andrea, Cook, Jonathan P., Koszinowski, Ulrich and Burgert, Hans-Gerhard (2006) Transposon-assisted cloning and traceless mutagenesis of adenoviruses: Development of a novel vector based on species D. Journal of Virology, Vol.80 (No.16). pp. 8100-8113. doi:10.1128/JVI.00687-06

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Official URL: http://dx.doi.org/10.1128/JVI.00687-06

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Abstract

Until recently, adenovirus (Ad)-mediated gene therapy was almost exclusively based on human Ad type 5 (Ad5). Preexisting immunity and the limited, coxsackievirus and adenovirus receptor-dependent tropism of Ad5 stimulated attempts to exploit the natural diversity in tropism of the other 50 known human Ad serotypes. Aiming in particular at immunotherapy and vaccination, we have screened representative serotypes from different Ad species for their ability to infect dendritic cells. Ad19a, an Ad from species D, was selected for development as a new vector for vaccination and cancer gene therapy. To clone and manipulate its genome, we have developed a novel methodology, coined "exposon mutagenesis," that allows the rapid and precise introduction of virtually any genetic alteration (deletions, point mutations, or insertions) into recombinant Ad bacterial artificial chromosomes. The versatility of the system was exemplified by deleting the E3 region of Ad19a, by specifically knocking out expression of a species-specific E3 gene, E3/49K, and by reinserting E3/49K into an E3 null Ad19a mutant. The technology requires only limited sequence information and is applicable to other Ad species. Therefore, it should be extremely valuable for the analysis of gene functions from any Ad species. In addition, a basic, replication-defective E1- and E3-deleted Ad19a vector expressing GFP (Ad19aGFP) was generated. This new vector based on species D Ads exhibits a very promising tropism for lymphoid and muscle cells and shows great potential as an alternative vector for transduction of cell types that are resistant to or only poorly transduced by conventional Ad5-based vectors.

Item Type: Journal Article
Subjects: Q Science > QR Microbiology > QR355 Virology
Divisions: Faculty of Science > Life Sciences (2010- ) > Biological Sciences ( -2010)
Faculty of Science > Life Sciences (2010- )
Journal or Publication Title: Journal of Virology
Publisher: American Society for Microbiology
ISSN: 0022-538X
Official Date: August 2006
Dates:
DateEvent
August 2006Published
Volume: Vol.80
Number: No.16
Number of Pages: 14
Page Range: pp. 8100-8113
DOI: 10.1128/JVI.00687-06
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Restricted or Subscription Access
Funder: Mercia Spinner Pathfinder, Deutsche Forschungsgemeinschaft (DFG), Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC)
Grant number: BU 642-1 and SFB455 (DFG), BB/D002877/1 (BBSRC)

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