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  • Nonviral Transfer Technology
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Cell cycle dependence of gene transfer by lipoplex, polyplex and recombinant adenovirus

Gene Therapy volume 7, pages 401–407 (2000)Cite this article

Abstract

The aim of this study was to investigate the influence of cell cycle on transfection efficiency. Counterflow centrifugal elutriation was used which avoids possible side-effects from chemical treatment of cells. With this method, cell populations were fractionated by means of size and density, and fractions corresponding to discrete cell cycle phase-specific populations were transfected with various nonviral methods (Lipofectamine, TfpLys and TfPEI), adenovirus-enhanced transferrinfection (AVET system) and recombinant adenovirus. Transfection efficiency was found to be strongly dependent on the cell cycle stage at the time of transfection. Luciferase activity from cells transfected with polycation- or lipid-based transfection systems was 30- to more than 500-fold higher when transfection was performed during S or G2 phase compared with cells in G1 phase which have the lowest expression levels. In contrast, this effect was not observed with recombinant adenovirus which varied only four-fold. Our results indicate that mitotic activity enhances transfection not only by lipoplexes but also by polyplexes, but not a viral system which has an efficient nuclear entry machinery, suggesting that transfection close to M phase is facilitated perhaps by nuclear membrane breakdown. Furthermore, low transfection success into G1 cells indicates that DNA complexes deposited in G1 cells are probably not retained long enough to take advantage of mitosis effects or that passage of transfected cells through S phase is inhibitory.

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Acknowledgements

We would like to thank Josef Gotzmann and Wolfgang Mikulits (Institut für Tumorbiologie und Krebsforschung) for the stably transfected HeLa cells. We thank Ingrid Mudrak for the HeLa cells that were elutriated and Karin Paiha and Peter Steinlein for help with FACS analysis. Thanks to Bettina Grosse, Helga Vetr, Thomas Blessing and Peter Wallner for inspiring discussions and for critically reading the manuscript. This work was supported by grant S07405 from the Austrian Fonds zur Förderung der Wissenschaftlichen Forschung.

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Authors and Affiliations

  1. Institute of Biochemistry, Unversity of Vienna, Vienna, Austria

    S Brunner, S Carotta & E Wagner

  2. Institute of Molecular Biology, Unversity of Vienna, Vienna, Austria

    T Sauer

  3. Institute for Molecular Pathology, Vienna, Austria

    M Cotten & M Saltik

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  1. S Brunner
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  2. T Sauer
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  3. S Carotta
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  4. M Cotten
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  5. M Saltik
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  6. E Wagner
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Brunner, S., Sauer, T., Carotta, S. et al. Cell cycle dependence of gene transfer by lipoplex, polyplex and recombinant adenovirus. Gene Ther 7, 401–407 (2000). https://doi.org/10.1038/sj.gt.3301102

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  • DOI: https://doi.org/10.1038/sj.gt.3301102

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