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1

The most important features of adenoviral vectors

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Stopa M., Dulak J., Jozkowicz A.
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issue 3
22-32
EN
One of the major hurdles to successful gene therapy is the ability to efficiently introduce a foreign nucleic acid into the tissue of interest. As adenoviruses posses ability to enter rapidly a mammalian host cell and achieve propagation, they are widely used as a tool for delivery transgenes. Recombinant adenoviral vectors of first generation (AdV) contain an expression cassette with exogenous genes and are made replication deficient by the deletion of the E1/E3 region. They offer many advantages for gene delivery: ability to transduce a wide variety of cell types in a cell-cycle independent manner, easy and cheap propagation process to high titers and low pathogenicity for humans. However, AdV also have some disadvantages, namely cytotoxity, immunogenity, transient expression of transgene, which are mostly important in case of clinical trials. Despite those limitations and development of more sophisticated adenoviral systems (helper-dependent adenoviral vectors), AdV of first generation are still widely used for trasnducting different cell types in vitro, especially those that are refractory to other gene transfer methods, as well as in gene therapy clinical trials.
2

Production of adenoviral vectors of first generation ? optimization of method

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Stopa M., Jozwa A., Mleczko J., Dulak A., Jozkowicz A.
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EN
The paper presents the production of adenoviral vectors (AdV) containing beta-galactosidase (Adbeta-gal), from the transfer of recombinant viral DNA into packing cell line (HEK293) to the titration of viral particles. Optimisation of the methods (preparation of DNA for transfection, adsorption time during the infection of cells, amount of serum in the medium, time-point of vector isolation) enables obtaining a titer of up to 1010 IU/mL. Adbeta-gal were titrated with several methods, with beta-gal in situ staining used as a reference. We found that the most suitable titration method of the vectors containing other than reporter genes was the Rapid Titer ELISA kit?.
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