Journal article

Generation of rho(0) cells utilizing a mitochondrially targeted restriction endonuclease and comparative analyses

Alexandra Kukat, Christian Kukat, Jan Brocher, Ingo Schaefer, Georg Krohne, Ian A Trounce, Gaetano Villani, Peter Seibel

NUCLEIC ACIDS RESEARCH | OXFORD UNIV PRESS | Published : 2008

Abstract

Eukaryotic cells devoid of mitochondrial DNA (rho0 cells) were originally generated under artificial growth conditions utilizing ethidium bromide. The chemical is known to intercalate preferentially with the mitochondrial double-stranded DNA thereby interfering with enzymes of the replication machinery. Rho0 cell lines are highly valuable tools to study human mitochondrial disorders because they can be utilized in cytoplasmic transfer experiments. However, mutagenic effects of ethidium bromide onto the nuclear DNA cannot be excluded. To foreclose this mutagenic character during the development of rho0 cell lines, we developed an extremely mild, reliable and timesaving method to generate rho0..

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University of Melbourne Researchers