Molecular Biotechnology

, Volume 34, Issue 1, pp 51–54 | Cite as

Enhanced DNA transfer into fish bone cells using polyethylenimine

  • Daniel Braga
  • Vincent Laizé
  • Daniel M. Tiago
  • M. Leonor Cancela
Research Protocols


The use of in vitro cell culture systems to assess gene function largely depends on the successful transfer of DNA into target cells. Well developed in mammals, transfection methods are still to be optimized for non-mammalian cell culture systems, like fish. Here we describe a rapid, cost-efficient, and successful method to transfer DNA into a fish bone-derived cell line using polyethylenimine (PEI) as the DNA carrier. Using this method, DNA transfer was remarkably enhanced in comparison with commercially available reagents, as demonstrated by the increased activity of both luciferase and green fluorescent protein observed in the transfected cells. Its efficiency in transferring DNA intoa wide range of cell types, including non-mammalian and hard-to-transfect cells, in addition to a low cost, show that PEI is a reagent of choice for nonviral vector transfection.

Index entries

DNA transfer cell transfection polyethylenimine (PEI) non-mammalian cells 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Du, S. L. and Haga, Y. (2004) The Zebrafish as a model for studying skeletal development, in Biomineralization: Progress in Biology, Molecular Bbiology and Application (Baeuerlein, E., ed.), Willey-VCH, Weinheim, Germany, pp. 283–304.Google Scholar
  2. 2.
    Inohaya, K. and Kudo, A. (2000) Medaka as a model organism of skeletal development. Tanpakushitsu Kakusan Koso 45, 2745–2751.PubMedGoogle Scholar
  3. 3.
    Pombinho, A. R., Laizé, V., Molha, D. M., Marques, S. M. P., and Cancela, M. L. (2004) Development of two bone-derived cell lines from the marine teleost Sparus aurata: evidence for extracellular matrix mineralization and cell-type-specific expression of matrix Gla protein and osteocalcin. Cell Tissue Res. 315, 393–406.PubMedCrossRefGoogle Scholar
  4. 4.
    Clamme, J. P., Azoulay, J., and Mely, Y. (2003) Monitoring the formation and dissociation of Polyethylenimine/DNA complexes by two proton fluorescence correlation spectroscopy. Biophys. J. 84, 1960–1968.PubMedCrossRefGoogle Scholar
  5. 5.
    Kunath, K., Harpe, A., Fisher, D., et al. (2003) Lowmolecular-weight polyethylenimine as a non-viral vector for DNA delivery: Comparison of physicochemical properties, transfection efficiency and in vivo distribution with high-molecular-weight polyethylenimine. J. Control. Release 89, 113–125.PubMedCrossRefGoogle Scholar
  6. 6.
    Allo, J. C., Midoux, P., Merten, M., et al. (2000) Efficient gene transfer into human normal and cystic fibrosis tracheal gland serous cells with synthetic vectors. Am. J. Respir. Cell Mol. Biol. 22, 166–175.PubMedGoogle Scholar
  7. 7.
    Suh, J., Wirtz, D., and Hanes, J. (2003) Efficient active transport of gene nanocarriers to the cell nucleus. Proc. Natl. Acad. Sci. U.S.A. 100, 3878–3882.PubMedCrossRefGoogle Scholar
  8. 8.
    Florea, B. I., Meaney, C., Junginger, H. E., and Borchard, G. (2002) Transfection efficiency and toxicity of polyethylenimine in differentiated Calu-3 and non-differentiated COS-1 cell cultures. AAPS PharmSci. 4, E12.PubMedCrossRefGoogle Scholar
  9. 9.
    Boussif, O., Lezoualc'h, F., Zanta, M. A., et al. (1995) A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: Polyethylenimine. Proc. Natl. Acad. Sci. U.S.A. 92, 7297–7301.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc 2006

Authors and Affiliations

  • Daniel Braga
    • 1
  • Vincent Laizé
    • 1
  • Daniel M. Tiago
    • 1
  • M. Leonor Cancela
    • 1
  1. 1.Centre of Marine Sciences (CCMAR)University of AlgarveFaroPortugal

Personalised recommendations