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Expression of GFP in the mitochondrial compartment using DQAsome-mediated delivery of an artificial mini-mitochondrial genome

  • Diana Lyrawati*
  • , Alan Trounson
  • , David Cram
  • *Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)2848-2862
Number of pages15
JournalPharmaceutical Research
Volume28
Issue number11
DOIs
Publication statusPublished - Nov 2011

Keywords

  • DQAsome
  • GFP expression
  • Minimitochondrial genome
  • Mitochondrial gene delivery
  • Transfection

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