Selection by drug resistance proteins located in the mitochondria of mammalian cells

Young Geol Yoon; Michael D. Koob

doi:10.1016/j.mito.2008.07.004

Selection by drug resistance proteins located in the mitochondria of mammalian cells

Young Geol Yoon, Michael D. Koob

Laboratory Medicine and Pathology

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	345-351
Number of pages	7
Journal	Mitochondrion
Volume	8
Issue number	5-6
DOIs	https://doi.org/10.1016/j.mito.2008.07.004
State	Published - Dec 2008

Bibliographical note

Funding 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).

Keywords

Drug-resistance proteins
Mitochondria
Selectable markers
mtDNA

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10.1016/j.mito.2008.07.004

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title = "Selection by drug resistance proteins located in the mitochondria of mammalian cells",

abstract = "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.",

keywords = "Drug-resistance proteins, Mitochondria, Selectable markers, mtDNA",

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note = "Funding 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).",

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AU - Yoon, Young Geol

AU - Koob, Michael D.

N1 - Funding 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).

PY - 2008/12

Y1 - 2008/12

N2 - 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.

AB - 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.

KW - Drug-resistance proteins

KW - Mitochondria

KW - Selectable markers

KW - mtDNA

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Selection by drug resistance proteins located in the mitochondria of mammalian cells

Abstract

Bibliographical note

Keywords

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