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Expression of GFP in the Mitochondrial Compartment Using DQAsome-Mediated Delivery of an Artificial Mini-mitochondrial Genome

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ABSTRACT

Purpose

We describe a novel strategy for expression of GFP in mammalian mitochondria.

Methods

The key components of the strategy were an artificially created mitochondrial genome pmtGFP and a DQAsome transfection system.

Results

Using immunofluorescence and a combination of immunohistochemical and molecular based techniques, we show that DQAsomes are capable of delivering the pmtGFP construct to the mitochondrial compartment of the mouse macrophage cell line RAW264.7, albeit at low efficiency (1–5%), resulting in the expression of GFP mRNA and protein. Similar transfection efficiencies were also demonstrated in a range of other mammalian cell lines.

Conclusions

The DQAsome-transfection technique was able to deliver the exogenous DNA into the cellular mitochondria and the pmtGFP was functional. Further optimization of this strategy would provide a flexible and rapid way to generate mutant cells and useful animal models of mitochondrial disease.

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ACKNOWLEDGMENTS

Dr Diana Lyrawati was supported by a Departmental Scholarship provided by the Monash Immunology and Stem Cell Laboratories, formerly, the Centre for Early Human Development, under the Directorship of Professor Alan Trounson. The authors thank Professor Sangkot Marzuki from the Eijkman Institute for Molecular Biology, Jakarta, Indonesia for providing laboratory resources to perform the immunohistochemistry experiments.

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Author notes

  1. Diana Lyrawati

    Present address: Laboratory of Pharmacy, Faculty of Medicine, Brawijaya University, Jl. Veteran, Malang, Indonesia

Authors and Affiliations

  1. California Institute for Regenerative Medicine, San Francisco, California, 94107, USA

    Alan Trounson

  2. Monash Immunology and Stem Cell Laboratories, Monash University, Melbourne, Australia

    Diana Lyrawati, Alan Trounson & David Cram

Authors
  1. Diana Lyrawati
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  2. Alan Trounson
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  3. David Cram
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Correspondence to Diana Lyrawati.

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Lyrawati, D., Trounson, A. & Cram, D. Expression of GFP in the Mitochondrial Compartment Using DQAsome-Mediated Delivery of an Artificial Mini-mitochondrial Genome. Pharm Res 28, 2848–2862 (2011). https://doi.org/10.1007/s11095-011-0544-0

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  • DOI: https://doi.org/10.1007/s11095-011-0544-0

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