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Polyethylenimine-mediated cellular uptake, nucleus trafficking and expression of cytokine plasmid DNA

Gene Therapy volume 9, pages 1627–1632 (2002)Cite this article

Abstract

Although polyethylenimine (PEI) has been widely used as a nonviral vector, there is little mechanistic understanding on PEI-mediated delivery. Here, we studied whether the expression of murine interleukin-2 (mIL-2) plasmids could be improved by complexation with PEI at various N/P ratios, and whether the cellular uptake, nuclear translocation, and retention of plasmids could be affected by the N/P ratios. Compared with the naked mIL-2, PEI/mIL-2 complexes showed at least two orders of magnitude higher expression at Raw264 cells in the N/P ratio-dependent manner. PEI-mediated cellular uptake and nuclear trafficking of plasmids, quantitated by competitive polymerase chain reaction, also depended on the N/P ratios showing the highest cell and nuclear levels of plasmids at 10/1. The higher cellular levels of plasmid DNA after PEI-mediated delivery were also observed in other cell lines. Unlike naked plasmids, PEI/mIL-2 complexes (N/P ratios ≥4/1) showed prolonged cellular and nuclear retention of mIL-2 plasmids. The nuclear translocation and higher cellular level of plasmids given in PEI complexes were similarly observed by fluorescence microscopy. Moreover, PEI/mIL-2 complexes revealed high stability against DNase I, partly explaining the prolonged subcellular retention. These results indicate that the expression of plasmid mIL-2 might be highly enhanced by complexation with PEI and that such increased expression could be attributed by the higher cellular uptake, nuclear translocation and prolonged retention.

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Acknowledgements

This work was supported by a grant of the Korea Research Foundation, Republic of Korea (KRF-99-041-F00110). We greatly appreciate Dr Young-Chul Sung (Pohang University of Science and Technology, Pohang, South Korea) for providing the murine IL-2 gene, pCIneoIL-2.

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Authors and Affiliations

  1. Department of Microbiology and Institute of Medical Research, College of Medicine, Pochon CHA University, Kyonggi-do, South Korea

    Y-K Oh, K Shin & J J Ko

  2. Department of Biochemistry, College of Medicine, Pochon CHA University, Kyonggi-do, South Korea

    D Suh

  3. Department of Microbiology and Institute of Biomedical Research, College of Medicine, Hanyang University, Seoul, South Korea

    J M Kim

  4. Department of Pharmaceutics, College of Pharmacy, Yeungnam University, Gyongsan, South Korea

    H-G Choi

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Oh, YK., Suh, D., Kim, J. et al. Polyethylenimine-mediated cellular uptake, nucleus trafficking and expression of cytokine plasmid DNA. Gene Ther 9, 1627–1632 (2002). https://doi.org/10.1038/sj.gt.3301735

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