Methods for efficient elimination of mitochondrial DNA from cultured cells
© 2016 Spadafora et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Here, we document that persistent mitochondria DNA (mtDNA) damage due to mitochondrial overexpression of the Y147A mutant uracil-N-glycosylase as well as mitochondrial overexpression of bacterial Exonuclease III or Herpes Simplex Virus protein UL12.5M185 can induce a complete loss of mtDNA (ρ0 phenotype) without compromising the viability of cells cultured in media supplemented with uridine and pyruvate. Furthermore, we use these observations to develop rapid, sequence-independent methods for the elimination of mtDNA, and demonstrate utility of these methods for generating ρ0 cells of human, mouse and rat origin. We also demonstrate that ρ0 cells generated by each of these three methods can serve as recipients of mtDNA in fusions with enucleated cells.
Publication Source (Journal or Book title)
Spadafora, D., Kozhukhar, N., Chouljenko, V., Kousoulas, K., & Alexeyev, M. (2016). Methods for efficient elimination of mitochondrial DNA from cultured cells. PLoS ONE, 11 (5) https://doi.org/10.1371/journal.pone.0154684