Selection by drug resistance proteins located in the mitochondria of mammalian cells.
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.
Yoon
Young Geol
YG
Department of Laboratory Medicine and Pathology, University of Minnesota, 420 Delaware Street S.E., Minneapolis, MN 55455, USA.
Koob
Michael D
MD
D049109:Cell Proliferation; D002934:Cinnamates / Q000494:pharmacology; D004351:Drug Resistance; D005839:Gentamicins / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D019868:Kanamycin Kinase / Q000096:biosynthesis*; D008928:Mitochondria / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000096:biosynthesis*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis
D002934:Cinnamates; D005839:Gentamicins; D011993:Recombinant Fusion Proteins; D006921:Hygromycin B; C026273:hygromycin A; C010680:antibiotic G 418; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase; D019868:Kanamycin Kinase