Release of Plasmid-DNC from PEI/DNA Particles

  • Martin Bertschinger
  • Arnaud Schertenleib
  • Gaurav Backliwal
  • Martin Jordan
  • Florian M. Wurm
Conference paper

Abstract

The cationic polymer polyethylenimine (PEI) spontaneously forms complexes with DNA that efficiently transfect mammalian cells. To study the reversibility of the DNA/PEI interaction in vitro, preformed complexes were diluted with varying amounts of plasmid DNA and RNA. The released plasmid DNA was visualized by gel electrophoresis. With this simple assay we determined the kinetics of the release of plasmid DNA. Even at very low concentrations RNA instantaneously replaced most of the plasmid DNA in the complex, whereas DNA-mediated release takes up to one day. The results of these in vitro experiments are important for understanding the transport and release of plasmid DNA from PEI/DNA particles after transfection of cells.

Keywords

Agarose Electrophoresis 

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References

  1. Bertschinger, M., Chaboche, S., Jordan, M., Wurm, FM. 2004. A spectrophotometric assay for the quantification of polyethylenimine in DNA nanoparticles. Anal Biochem. 334: 196–8.CrossRefGoogle Scholar
  2. Derouazi, M., Girard, P., Van Tilborgh, F., Iglesias, K., Muller, N., Bertschinger, M., Wurm FM. 2004. Serum-free large-scale transient transfection of CHO cells. Biotechnol Bioeng. 87: 537–45.CrossRefGoogle Scholar
  3. Hacker, DL., Bertschinger, M., Baldi, L., Wurm, FM. 2004 Reduction of adenovirus E1A mRNA by RNAi results in enhanced recombinant protein expression in transiently transfected HEK293 cells. Gene 341: 227–34.CrossRefGoogle Scholar

Copyright information

© Springer 2007

Authors and Affiliations

  • Martin Bertschinger
    • 1
  • Arnaud Schertenleib
    • 1
  • Gaurav Backliwal
    • 1
  • Martin Jordan
    • 1
  • Florian M. Wurm
    • 1
  1. 1.Laboratory of Cellular BiotechnologyEcole Polytechnique Federale de Lausanne (EPFL)LausanneSwitzerland

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