Establishment of human cell lines lacking mitochondrial DNA

Methods Mol Biol. 2009;554:383-91. doi: 10.1007/978-1-59745-521-3_23.

Abstract

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 rho(0) cells) are very effective tools for studying the consequences of mtDNA mutations. rho(0)cell 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 rho(0) cell lines that have been studied. We provide simple protocols for the generation of human rho(0) cells by exposure to ethidium bromide and PCR verification of their rho(0) status.

MeSH terms

  • Cell Culture Techniques / methods*
  • Cells, Cultured
  • DNA, Mitochondrial / genetics*
  • Ethidium / pharmacology
  • Humans
  • Indicators and Reagents / pharmacology
  • Mitochondria / drug effects*
  • Mitochondria / metabolism*
  • Polymerase Chain Reaction / methods*

Substances

  • DNA, Mitochondrial
  • Indicators and Reagents
  • Ethidium