Manipulation of mitochondrial genes and mtDNA heteroplasmy

Methods Cell Biol. 2020;155:441-487. doi: 10.1016/bs.mcb.2019.12.004. Epub 2020 Jan 20.


Most patients with mitochondrial DNA (mtDNA) mutations have a mixture of mutant and wild-type mtDNA in their cells. This phenomenon, known as mtDNA heteroplasmy, provides an opportunity to develop therapies by selectively eliminating the mutant fraction. In the last decade, several enzyme-based gene editing platforms were developed to cleave specific DNA sequences. We have taken advantage of these enzymes to develop reagents to selectively eliminate mutant mtDNA. The replication of intact mitochondrial genomes normalizes mtDNA levels and consequently mitochondrial function. In this chapter, we describe the methodology used to design and express these nucleases in mammalian cells in culture and in vivo.

Keywords: Gene therapy; Heteroplasmy; Mitochondrial diseases; mitoTALEN; mtDNA; mtZFN.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COS Cells
  • Chlorocebus aethiops
  • DNA, Mitochondrial / genetics*
  • Female
  • Genes, Mitochondrial*
  • HeLa Cells
  • Heteroplasmy / genetics*
  • Humans
  • Mice
  • Mutation / genetics
  • Plasmids / genetics
  • Transcription Activator-Like Effector Nucleases
  • Zinc Finger Nucleases / metabolism


  • DNA, Mitochondrial
  • Transcription Activator-Like Effector Nucleases
  • Zinc Finger Nucleases