Transfection of mitochondria: strategy towards a gene therapy of mitochondrial DNA diseases

Nucleic Acids Res. 1995 Jan 11;23(1):10-7. doi: 10.1093/nar/23.1.10.


Successes in classical gene therapies have been achieved by placing a corrected copy of a defective nuclear gene in cells. A similar gene replacement approach for a mutant mitochondrial genome is invariably linked to the use of a yet unavailable mitochondrial transfection vector. Here we show that DNA coupled covalently to a short mitochondrial leader peptide (chimera) can enter mitochondria via the protein import pathway, opening a new way for gene-, antisense-RNA- or antisense-DNA-delivery in molecular therapies. The import behavior of the purified chimera, composed of the amino-terminal leader peptide of the rat ornithine transcarbamylase (OTC) and a double stranded DNA molecule (17 bp or 322 bp), was tested by incubating with coupled and 'energized' rat liver mitochondria in the presence of reticulocyte lysate. The chimera was translocated with a high efficiency into the matrix of mitochondria utilizing the protein import pathway, independent from the size of its passenger DNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Biological Transport, Active
  • DNA, Mitochondrial / genetics*
  • DNA, Mitochondrial / metabolism
  • Genetic Therapy*
  • In Vitro Techniques
  • Mitochondria, Liver / metabolism*
  • Molecular Sequence Data
  • Mutation
  • Ornithine Carbamoyltransferase / genetics
  • Ornithine Carbamoyltransferase / metabolism
  • Protein Sorting Signals / genetics
  • Protein Sorting Signals / metabolism
  • Rats
  • Rats, Wistar
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Transfection*


  • DNA, Mitochondrial
  • Protein Sorting Signals
  • Recombinant Fusion Proteins
  • Ornithine Carbamoyltransferase