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Enhanced cationic liposome-mediated transfection using the DNA-binding peptide μ (mu) from the adenovirus core

Gene Therapy volume 8, pages 453–460 (2001)Cite this article

Abstract

Promising advances in nonviral gene transfer have been made as a result of the production of cationic liposomes formulated with synthetic cationic lipids (cytofectins) that are able to transfect cells. However few cationic liposome systems have been examined for their ability to transfect CNS cells. Building upon our earlier use of cationic liposomes formulated from 3β-[N-(N′,N′-dimethylaminoethane)carbamoyl] cholesterol (DC-Chol) and dioleoyl-L-α-phosphatidyl-ethanolamine (DOPE), we describe studies using two cationic viral peptides, μ (mu) and Vp1, as potential enhancers for cationic liposome-mediated transfection. Mu is derived from the condensed core of the adenovirus and was selected to be a powerful nucleic acid charge neutralising and condensing agent. Vp1 derives from the polyomavirus and harbours a classical nuclear localisation signal (NLS). Vp1 proved disappointing but lipopolyplex mixtures formulated from pCMVβ plasmid, mu peptide and DC-Chol/DOPE cationic liposomes were able to transfect an undifferentiated neuronal ND7 cell line with β-galactosidase reporter gene five-fold more effectively than lipoplex mixtures prepared from pCMVβ plasmid and DC-Chol/DOPE cationic liposomes. Mu was found to give an identical enhancement to cationic liposome-mediated transfection of ND7 cells as poly-L-lysine (pLL) or protamine sulfate (PA). The enhancing effects of mu were found to be even greater (six- to 10-fold) when differentiated ND7 cells were transfected with mu-containing lipopolyplex mixtures. Differentiated ND7 cells represent a simple ex vivo-like post-mitotic CNS cell system. Successful transfection of these cells bodes well for transfection of primary neurons and CNS cells in vivo. These findings have implications for experimental and therapeutic uses of cationic liposome-mediated delivery of nucleic acids to CNS cells.

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Acknowledgements

This work was supported by a grant from the Motor Neurone Disease Association. SIS thanks the BBSRC and Genzyme for a CASE studentship. We thank the Mitsubishi-Tokyo Pharmaceutical company for supporting the Imperial College Genetic Therapies Centre and the West Riding Medical Research Trust for their support.

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Authors and Affiliations

  1. Department of Psychiatry and Behavioural Sciences, University College London Medical School, Windeyer Institute of Medical Sciences, London, UK

    K D Murray & H M D Gurling

  2. Department of Chemistry, Imperial College Genetic Therapies Centre, Imperial College of Science, Technology and Medicine, London, UK

    C J Etheridge, S I Shah & A D Miller

  3. Molecular Medicine Unit, University of Leeds, St James University Hospital, Leeds, UK

    D A Matthews

  4. School of Biomedical Sciences, University of St Andrews, UK

    W Russell

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  1. K D Murray
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Murray, K., Etheridge, C., Shah, S. et al. Enhanced cationic liposome-mediated transfection using the DNA-binding peptide μ (mu) from the adenovirus core. Gene Ther 8, 453–460 (2001). https://doi.org/10.1038/sj.gt.3301401

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