Transgene amplification and persistence after delivery of retroviral vector and packaging functions with E1/E4-deleted adenoviruses View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2000-08

AUTHORS

Christophe Torrent, Carole Jullien, David Klatzmann, Michel Perricaudet, Patrice Yeh

ABSTRACT

Adenovirus DNA is rapidly lost in actively dividing cells. In addition, first-generation (E1-defective) vectors trigger a strong cytotoxicity that impairs the duration of transgene expression. To solve these issues, we have developed a chimeric vector system that uses E1/E4 doubly defective adenoviruses for efficient production of infectious retroviral vectors. The retroviral vector sequences and packaging functions were split into two E1/E3/E4-deleted adenoviral vectors: the Moloney murine leukemia virus gag-pol cistron was expressed from the human EF1 alpha (elongation factor) promoter (AdGAG/POL), whereas the thymidine kinase transgene, embedded in a retroviral vector context, and an amphotropic retroviral envelope cassette were included within a second adenovirus (AdTK/ENV). This chimeric vector system was evaluated with a special emphasis on recombinant retrovirus production in vitro, as well as transgene amplification and persistence in vivo. Retrovirus titers of >10(5) infectious units/mL were routinely obtained in W162 cells coinfected with both recombinant adenoviruses. Long-term transgene persistence (up to 3 months) was demonstrated in vitro in two different cell lines coinfected with AdGAG/POL and AdTK/ENV, and correlated with the detection of specific provirus sequences. A 10- to 50-fold transgene amplification also was demonstrated in an in vivo tumor model infected with the Ad/Rt chimeric vector system. The chimeric vector system described herein combines the efficiency of gene delivery by recombinant adenoviruses with the integrative properties of infectious retroviral vectors. This versatile vector system may open up new avenues for efficient production of oncogenic, but also non-oncogenic, retroviruses from cells of non-murine origin. More... »

PAGES

1135

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/sj.cgt.7700212

DOI

http://dx.doi.org/10.1038/sj.cgt.7700212

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1032974917

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/10975674


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