2005/09/07 by Young Geol Yoon · 1 citation
Biochemistry, Genetics and Molecular Biology · #Mitochondrial Function and Pathology #ATP Synthase and ATPases Research #RNA and protein synthesis mechanisms
paper · pdf · doi:10.1093/nar/gni140
openalex publication_date 2005/09/07 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/30
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.