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Nonviral gene transfer into fetal mouse livers (a comparison between the cationic polymer PEI and naked DNA)

Gene Therapy volume 10, pages 810–817 (2003)Cite this article

Abstract

We investigated the efficacy and safety of the cationic polymer polyethylenimine (PEI) as a potential tool for intrauterine gene delivery into livers of fetal mice in the last trimester of pregnancy (E17.5). Using luciferase as a reporter gene, transferrin-conjugated and ligand-free PEI/DNA complexes (containing 3 μg DNA) with varying PEI-nitrogen/DNA-phosphate (N/P) ratios and different PEI forms, branched (800, 25 kDa) and linear (22 kDa), were compared with naked DNA. Transgene expression was measured 48 h after administration of PEI/DNA complexes or naked DNA. Highest luciferase activity (9.8 × 103 relative light units (RLU)/mg of tissue protein) was observed with ligand-free PEI22/DNA mixtures at N/P 6.0. In addition, this formulation was associated with very low toxicity as compared to the other PEI/DNA-injected groups. Using β-galactosidase as a reporter gene, transfection of single, but also small, clusters of cells was demonstrated throughout the liver. Injection of 3 μg naked DNA resulted in an 11-fold lower transgene expression value (0.9 × 103 RLU/mg of tissue protein) as compared to PEI22/DNA complexes. However, the administration of higher concentrated naked DNA (9 μg) into fetal livers yielded expression levels of 3.2 × 104 RLU/mg of tissue protein, a more than three-fold increase compared to PEI22/DNA complexes. Furthermore, the gene transfer efficacy of concentrated naked DNA was approximately 40 times higher in fetuses than in adults (0.8 × 103 RLU/mg of tissue protein), indicating that fetal tissue is especially amenable to the uptake and expression of naked DNA.

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Acknowledgements

We thank Andrea Fuchsbichler for performing electron microscopy, Vanessa Rössler for technical help with luciferase measurement and Dr Charles Buck for critically reading the manuscript. This work was supported by grants from the Austrian Science Fund (S 7401- MOB to KZ) and the Austrian Federal Ministry for Social Security and Generations (to KZ).

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Authors and Affiliations

  1. Institute of Pathology, University of Graz, Auenbruggerplatz, Graz, Styria, Austria

    H Gharwan & K Zatloukal

  2. Boehringer Ingelheim R&D Vienna, Dr Boehringer-Gasse, Vienna, Austria

    L Wightman & R Kircheis

  3. Pharmaceutical Biology – Biotechnology, Ludwig-Maximilians-University Munich, Munich, Germany

    E Wagner

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  1. H Gharwan
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  2. L Wightman
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Gharwan, H., Wightman, L., Kircheis, R. et al. Nonviral gene transfer into fetal mouse livers (a comparison between the cationic polymer PEI and naked DNA). Gene Ther 10, 810–817 (2003). https://doi.org/10.1038/sj.gt.3301954

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