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Nonreplicating intracellular bacterial vector for conjugative DNA transfer into mitochondria
Young Geol Yoon1, Michael D Koob
1Department of Biomedical Sciences, Jungwon University, 5 Dongbu-ri, Goesan-eup, Goesan-gun, Chungcheongbuk-do, Korea, 367-805.
Pharmaceutical Research
|February 22, 2012
Summary
Researchers engineered an E. coli strain for intracellular DNA transfer to mammalian cells. This bacterial vector can invade cells and survive, but transferring DNA to mitochondria requires further investigation.
Area of Science:
- Microbiology
- Cell Biology
- Genetic Engineering
Background:
- Previous work demonstrated DNA transfer into isolated mitochondria via E. coli conjugation.
- Mammalian cell mitochondria are targets for genetic material delivery.
Purpose of the Study:
- To engineer a conjugative E. coli strain capable of invading mammalian cells.
- To facilitate DNA transfer into the cytoplasm and subsequently to mitochondria within mammalian cells.
Main Methods:
- Developed a method to generate nonreplicating E. coli daughter cells.
- Modified an enteroinvasive E. coli (EIEC) strain with conjugative functions.
- Utilized EIEC for invasion and survival within mammalian tissue culture cells.
Main Results:
- Successfully created E. coli strains producing nonreplicating cells for conjugation.
- Demonstrated invasion of mammalian cells by the engineered E. coli.
- Confirmed survival of E. coli within the host cell cytoplasm for several days without causing cell death.
Conclusions:
- An E. coli host suitable for intracellular conjugative transfer was constructed.
- The ability of these bacterial vectors to transfer DNA into intracellular mitochondria remains unconfirmed.
- Further development of mitochondrial screening and selectable markers is needed.
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