Transformation of mitochondria in mammalian cells is now a technical challenge. In this report, we demonstrate that the standard drug resistant genes encoding neomycin and hygromycin phosphotransferases can potentially be used as selectable markers for mammalian mitochondrial transformation. We re-engineered the drug resistance genes to express proteins targeted to the mitochondrial matrix and confirmed the location of the proteins in the cells by fusing them with GFP and by Western blot and mitochondrial content mixing analyses. We found that the mitochondrially targeted-drug resistance proteins confer resistance to high levels of G418 and hygromycin without affecting the viability of cells.
Bibliographical noteFunding Information:
We thank Mycosol Inc. for the gift of a mitochondria-specific dye, and the generous financial support from the Minnesota Medical Foundation, the Academic Health Center, the Institute of Human Genetics of the University of Minnesota and National Institute of Health (NINDS Grant No. NS052612).
Copyright 2009 Elsevier B.V., All rights reserved.
- Drug-resistance proteins
- Selectable markers