Establishment of Human Cell Lines Lacking Mitochondrial DNA

Mitochondria have their own genome, and mitochondrial DNA (mtDNA) encodes 2 ribosomal RNAs, 22 transfer RNAs, and 13 polypeptides that function in oxidative phosphorylation (OXPHOS). mtDNA mutations lead to dysfunction of OXPHOS, resulting in cell death and/or compromised cellular activity. Cell lines lacking mtDNA (termed ρ0 cells) are very effective tools for studying the consequences of mtDNA mutations. ρ0cell lines have been used widely to investigate relationships between mtDNA mutation, mitochondrial function, and a variety of cellular processes. In this chapter, we summarize the yeast and animal ρ0 cell lines that have been studied. We provide simple protocols for the generation of human ρ0 cells by exposure to ethidium bromide and PCR verification of their ρ0 status.