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Bactofection of mammalian cells by Listeria monocytogenes: improvement and mechanism of DNA delivery

Gene Therapy volume 10, pages 2036–2045 (2003)Cite this article

Abstract

Bacteria-mediated transfer of plasmid DNA into mammalian cells (bactofection) is a potent approach to express plasmid-encoded heterologous proteins (protein antigens, toxins or enzymes) in a large set of different cell types including phagocytic and nonphagocytic mammalian cells. Previously, we have described a Listeria monocytogenes-mediated DNA delivery system, which releases plasmid DNA directly into the cytosol of mammalian cells by partial self-destruction of the carrier bacteria. Here we report on a second generation of this phage lysin supported bactofection system, which is greatly improved with respect to plasmid stability, transfer efficacy and biosafety. In this case, DNA release is initiated by spontaneous bacterial lysis in the infected cells cytosol which is subsequently enhanced by the simultaneously released phage lysin produced by the intracellular carrier bacteria. Bacteria that are capable of cell-to-cell spread are found to be much more efficient in bactofection than their nonspreading counterparts.

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Acknowledgements

We thank M Kuhn and B Joseph for the critical reading of the manuscript and Ch Berberich for the kind gift of KMP-11 cDNA. This work was supported by a grant from the Deutsche Forschungsgemeinschaft (Go168/27-1) and the Fonds der Chemischen Industrie. SP thanks the DFG Graduate College (GK520), and CS The BMBF (IZKF Würzburg, 01KS9603) for financial support.

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Author notes

  1. S Pilgrim and J Stritzker: Both these authors contributed equally to this work

Authors and Affiliations

  1. Theodor-Boveri-Institut (Biozentrum) der Universität Würzburg, Lehrstuhl für Mikrobiologie, Am Hubland, Würzburg, Germany

    S Pilgrim, J Stritzker, C Schoen, A Kolb-Mäurer, I Gentschev & W Goebel

  2. Institut fur Medizinische Mikrobiologie und Hygiene, Fakultät für Klinische Medizin Mannheim der Universität Heidelberg, Mannheim, Germany

    G Geginat

  3. Institut für Lebensmittel- und Ernährungswissenschaften, ETH Zuerich, Zuerich, Switzerland

    M J Loessner

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  1. S Pilgrim
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  4. A Kolb-Mäurer
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Pilgrim, S., Stritzker, J., Schoen, C. et al. Bactofection of mammalian cells by Listeria monocytogenes: improvement and mechanism of DNA delivery. Gene Ther 10, 2036–2045 (2003). https://doi.org/10.1038/sj.gt.3302105

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