Absence of both MGME1 and POLG EXO abolishes mtDNA whereas absence of either creates unique mtDNA duplications

Gonzalez, Christian D.; Nissanka, Nadee; Van Booven, Derek; Griswold, Anthony J.; Moraes, Carlos T.

doi:10.1016/j.jbc.2024.107128

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2024

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Cited by 2 publications

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Enzymatic tools for mitochondrial genome manipulation

Rimskaya,

Shebanov,

Entelis

et al. 2024

Biochimie

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Enzymatic tools for mitochondrial genome manipulation

Rimskaya,

Shebanov,

Entelis

et al. 2024

Biochimie

View full text Add to dashboard Cite

Engineering mtDNA Deletions by Reconstituting End-Joining in Human Mitochondria

Fu,

Land,

Cui

et al. 2024

Preprint

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Recent breakthroughs in the genetic manipulation of mitochondrial DNA (mtDNA) have enabled the precise introduction of base substitutions and the effective removal of genomes carrying harmful mutations. However, the reconstitution of mtDNA deletions responsible for severe mitochondrial myopathies and age-related diseases has not yet been achieved in human cells. Here, we developed a method to engineer specific mtDNA deletions in human cells by co-expressing end-joining (EJ) machinery and targeted endonucleases. As a proof-of-concept, we used mito-EJ and mito-ScaI to generate a panel of clonal cell lines harboring a ~3.5 kb mtDNA deletion with the full spectrum of heteroplasmy. Investigating these isogenic cells revealed a critical threshold of ~75% deleted genomes, beyond which cells exhibited depletion of OXPHOS proteins, severe metabolic disruption, and impaired growth in galactose-containing media. Single-cell multiomic analysis revealed two distinct patterns of nuclear gene deregulation in response to mtDNA deletion accumulation; one triggered at the deletion threshold and another progressively responding to increasing heteroplasmy. In summary, the co-expression of mito-EJ and programable nucleases provides a powerful tool to model disease-associated mtDNA deletions in different cell types. Establishing a panel of cell lines with a large-scale deletion at varying levels of heteroplasmy is a valuable resource for understanding the impact of mtDNA deletions on diseases and guiding the development of potential therapeutic strategies.

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Absence of both MGME1 and POLG EXO abolishes mtDNA whereas absence of either creates unique mtDNA duplications

Cited by 2 publications

References 35 publications

Enzymatic tools for mitochondrial genome manipulation

Enzymatic tools for mitochondrial genome manipulation

Engineering mtDNA Deletions by Reconstituting End-Joining in Human Mitochondria

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