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Peptide nucleic acid delivery to human mitochondria

Gene Therapy volume 6, pages 1919–1928 (1999)Cite this article

A Corrigendum to this article was published on 15 May 2000

Abstract

Peptide nucleic acids (PNAs) are synthetic polynucleobase molecules, which bind to DNA and RNA with high affinity and specificity. Although PNAs have enormous potential as anti-sense agents, the success of PNA-mediated gene therapy will require efficient cellular uptake and sub-cellular trafficking. At present these mechanisms are poorly understood. To address this, we have studied the uptake of biotinylated PNAs into cultured cell lines using fluorescence confocal microscopy. In human myoblasts, initial punctate staining was followed by the release of PNAs into the cytosol and subsequent localisation and concentration in the nucleus. To determine whether PNAs could also be used as therapeutic agents for mtDNA disease, we attempted to localise PNAs to the mitochondrial matrix. When attached to the presequence peptide of the nuclear-encoded human cytochrome c oxidase (COX) subunit VIII, the biotinylated PNA was successfully imported into isolated organelles in vitro. Furthermore, delivery of the biotinylated peptide-PNA to mitochondria in intact cells was confirmed by confocal microscopy. These studies demonstrate that biotinylated PNAs can be directed across cell membranes and to a specific sub-cellular compartment within human cells – highlighting the importance of these novel molecules for human gene therapy.

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Acknowledgements

PFC is a Wellcome Trust Clinical Research Fellow. RWT is funded by the Muscular Dystrophy Campaign. We are very grateful to Dr ZMA Chrzanowska-Lightowlers for her help with the various cultured cell lines. KD and RL acknowledge funding by grants of the Sonderforschungsbereich 286 of Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie. DMT and RNL are indebted to the Muscular Dystrophy Campaign and the Wellcome Trust for continuing support.

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Author notes

  1. P F Chinnery and R W Taylor: The first two authors contributed equally to this work

Authors and Affiliations

  1. Department of Neurology, The University of Newcastle upon Tyne, UK

    P F Chinnery, R W Taylor, D M Turnbull & R N Lightowlers

  2. Institut für Zytobiologie der Philipps-Universität Marburg, Marburg, Germany

    K Diekert & R Lill

Authors
  1. P F Chinnery
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  2. R W Taylor
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  3. K Diekert
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  4. R Lill
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  5. D M Turnbull
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  6. R N Lightowlers
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Chinnery, P., Taylor, R., Diekert, K. et al. Peptide nucleic acid delivery to human mitochondria. Gene Ther 6, 1919–1928 (1999). https://doi.org/10.1038/sj.gt.3301061

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