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Development of a novel gene delivery scaffold utilizing colloidal gold–polyethylenimine conjugates for DNA condensation

Gene Therapy volume 10, pages 1882–1890 (2003)Cite this article

Abstract

We have developed a novel gene delivery scaffold based on DNA plasmid condensation with colloidal gold/polyethylenimine conjugates. This scaffold system was designed to enable systematic study of the relationships between DNA complex physical properties and transfection efficiency. Using an enhanced green fluorescent protein-coding reporter plasmid and a Chinese hamster ovary cell line, we have measured the transfection efficiencies of our complexes using flow cytometry and their cytotoxicities using the trypan blue assay. We have also assayed complex particle morphologies using atomic force microscopy, photon correlation spectroscopy, and a novel plasmon absorbance peak position analysis. We achieved comparable rates of transfection relative to the commonly used polycationic condensation agents calcium phosphate and LipofectAMINE™, with comparably low cytotoxicities. In addition, by manipulating colloidal gold concentration, we could partially decouple complex physical properties including charge ratio, size, DNA loading, and polyethylenimine concentration. Our morphological analyses showed that complexes with a diameter of a few hundred nanometers and a charge ratio of ∼8 perform best in our transfection efficiency assays. The use of colloidal gold as a component in our delivery system provides a versatile system for manipulating complex properties and morphology as well as a convenient scaffold for planned ligand conjugation studies.

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Acknowledgements

Partial support of this work was provided by the Department of Chemical Engineering at Carnegie Mellon University, a Clare Boothe Luce Fellowship to MS, and an ICI Summer Research Fellowship to JG. We also express our appreciation to Greg Fisher and Christoffer Lagerholm in the Center for Light Microscope Imaging and Biotechnology for his help with fluorescence microscopy and flow cytometry; to Carrie Doonan, Adam Linstedt, and Amy Mehta in the Department of Biology for their assistance with cell culturing; and to Ijeoma Nnebe in the Department of Chemical Engineering for her help with AFM.

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  1. Department of Chemical Engineering, Carnegie Mellon University, Pittsburgh, PA, USA

    M M Ow Sullivan, J J Green & T M Przybycien

  2. Department of Biomedical Engineering, Carnegie Mellon University, Pittsburgh, PA, USA

    T M Przybycien

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  1. M M Ow Sullivan
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Ow Sullivan, M., Green, J. & Przybycien, T. Development of a novel gene delivery scaffold utilizing colloidal gold–polyethylenimine conjugates for DNA condensation. Gene Ther 10, 1882–1890 (2003). https://doi.org/10.1038/sj.gt.3302083

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