Specific elimination of mutant mitochondrial genomes in patient-derived cells by mitoTALENs

Nature Medicine volume 19, pages1111–1113(2013)Cite this article

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Abstract

Mitochondrial diseases are commonly caused by mutated mitochondrial DNA (mtDNA), which in most cases coexists with wild-type mtDNA, resulting in mtDNA heteroplasmy. We have engineered transcription activator-like effector nucleases (TALENs) to localize to mitochondria and cleave different classes of pathogenic mtDNA mutations. Mitochondria-targeted TALEN (mitoTALEN) expression led to permanent reductions in deletion or point-mutant mtDNA in patient-derived cells, raising the possibility that these mitochondrial nucleases can be therapeutic for some mitochondrial diseases.

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Figure 1: Rationale, verification and efficacy of the Δ5-mitoTALEN for elimination of mtDNAs with the common deletion.
Figure 2: Rationale and efficacy of the 14459A-mitoTALEN against the pathogenic m.14459A mtDNA point mutation.

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Acknowledgements

This work was supported by US National Institutes of Health grants 5R01EY010804, 1R01AG036871 and 1R01NS079965, the Muscular Dystrophy Association and The JDM Fund. We thank M. Hirano (Columbia University) for the G14459A cells, F. Delacote (Cellectis bioresearch) for expert assistance in designing TALENs, G. Zhai for critically reading the manuscript and A. Pickrell for critical scientific suggestions.

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Author notes

  1. Sandra R Bacman and Siôn L Williams: These authors contributed equally to this work.

Affiliations

  1. Department of Neurology, University of Miami Miller School of Medicine, Miami, Florida, USA

    Sandra R Bacman, Siôn L Williams, Milena Pinto, Susana Peralta & Carlos T Moraes

  2. Department of Cell Biology, University of Miami Miller School of Medicine, Miami, Florida, USA

    Carlos T Moraes

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  1. Sandra R Bacman
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  2. Siôn L Williams
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  4. Susana Peralta
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Contributions

S.R.B. and S.L.W. designed and conducted most experiments and assisted in writing the manuscript. M.P. assisted in the quantification of mtDNA deletions and in writing the manuscript. S.P. conducted the complex I activity experiments and assisted in writing the manuscript. C.T.M. designed and supervised the project and wrote the manuscript.

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Correspondence to Carlos T Moraes.

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The authors declare no competing financial interests.

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Supplementary Figures 1–6 and Supplementary Table 1 (PDF 12499 kb)

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Bacman, S., Williams, S., Pinto, M. et al. Specific elimination of mutant mitochondrial genomes in patient-derived cells by mitoTALENs. Nat Med 19, 1111–1113 (2013). https://doi.org/10.1038/nm.3261

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