Transformation of isolated mammalian mitochondria by bacterial conjugation
Open Access
- 7 September 2005
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 33 (16) , e139
- https://doi.org/10.1093/nar/gni140
Abstract
We have developed a method for transferring exogenous DNA molecules into isolated mammalian mitochondria using bacterial conjugation. In general, we accomplish this by (i) inserting an origin of DNA transfer (oriT) sequence into a DNA construct, (ii) transforming the construct into an appropriate Escherichia coli strain and then (iii) introducing the mobilizable DNA into mitochondria through conjugation. We tested this approach by transferring plasmid DNA containing a T7 promoter sequence into mitochondria that we had engineered to contain T7 RNA polymerase. After conjugation between E.coli and mitochondria, we detected robust levels of T7 transcription from the DNA constructs that had been transferred into the mitochondria. This approach for engineering DNA constructs in vitro and subsequent transfer into mitochondria by conjugation offers an attractive experimental system for studying many aspects of vertebrate mitochondrial gene expression and is a potential route for transforming mitochondrial networks within mammalian cells.Keywords
This publication has 29 references indexed in Scilit:
- Mitochondria Transfection by Oligonucleotides Containing a Signal Peptide and Vectorized by Cationic LiposomesAntisense and Nucleic Acid Drug Development, 2001
- Peptide nucleic acid delivery to human mitochondriaGene Therapy, 1999
- Mitochondrial EvolutionScience, 1999
- Conjugative transfer in the dissemination of beta-lactam and aminoglycoside resistanceFrontiers in Bioscience-Landmark, 1999
- Trans-kingdom conjugation offers a powerful gene targetingGenetic Analysis: Biomolecular Engineering, 1998
- A single mouse gene encodes the mitochondrial transcription factor A and a testis–specific nuclear HMG-box proteinNature Genetics, 1996
- Minimizing the Genome of Escherichia coliAnnals of the New York Academy of Sciences, 1994
- Hyper‐invasive mutants define a novel Pho‐regulated invasion pathway in Escherichia coliMolecular Microbiology, 1993
- Kinetic analysis of T7 RNA polymerase transcription initiation from promoters containing single-stranded regionsBiochemistry, 1993
- DNA-protein conjugates can enter mitochondria via the protein import pathwayNature, 1989