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A Mitochondrial Targeted Meganuclease Eliminates Mutant Mitochondrial DNA in a Mouse Model

Lookup NU author(s): Dr Jim StewartORCiD

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

Diseases caused by heteroplasmic mitochondrial DNA mutations have no effective treatment or cure. In recent years, DNA editing enzymes were tested as tools to eliminate mutant mtDNA in heteroplasmic cells and tissues. Mitochondrial-targeted restriction endonucleases, ZFNs, and TALENs have been successful in shifting mtDNA heteroplasmy, but they all have drawbacks as gene therapy reagents, including: large size, heterodimeric nature, inability to distinguish single base changes, or low flexibility and effectiveness. Here we report the adaptation of a gene editing platform based on the I-CreI meganuclease known as ARCUS®. These mitochondrial-targeted meganucleases (mitoARCUS) have a relatively small size, are monomeric, and can recognize sequences differing by as little as one base pair. We show the development of a mitoARCUS specific for the mouse m.5024C>T mutation in the mt-tRNAAla gene and its delivery to mice intravenously using AAV9 as a vector. Liver and skeletal muscle show robust elimination of mutant mtDNA with concomitant restoration of mt-tRNAAla levels. We conclude that mitoARCUS is a potential powerful tool for the elimination of mutant mtDNA.


Publication metadata

Author(s): Zekonyte U, Bacman S, Smith J, Sharer W, Tomberlin G, Stewart JB, Jantz D, Moraes C

Publication type: Article

Publication status: Published

Journal: Nature Communications

Year: 2021

Volume: 12

Online publication date: 28/05/2021

Acceptance date: 26/04/2021

Date deposited: 31/05/2021

ISSN (electronic): 2041-1723

Publisher: Springer Nature

URL: https://doi.org/10.1038/s41467-021-23561-7

DOI: 10.1038/s41467-021-23561-7


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Funding

Funder referenceFunder name
1R01AG036871
1R01NS079965
5R01EY010804

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