mitoTev‐TALE: a monomeric DNA editing enzyme to reduce mutant mitochondrial DNA levels
Abstract Pathogenic mitochondrial DNA (mtDNA) mutations often co‐exist with wild‐type molecules (mtDNA heteroplasmy). Phenotypes manifest when the percentage of mutant mtDNA is high (70–90%). Previously, our laboratory showed that mitochondria‐targeted transcription activator‐like effector nucleases...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Springer Nature
2018-09-01
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| Series: | EMBO Molecular Medicine |
| Subjects: | |
| Online Access: | https://doi.org/10.15252/emmm.201708084 |
| _version_ | 1797283867754758144 |
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| author | Claudia V Pereira Sandra R Bacman Tania Arguello Ugne Zekonyte Sion L Williams David R Edgell Carlos T Moraes |
| author_facet | Claudia V Pereira Sandra R Bacman Tania Arguello Ugne Zekonyte Sion L Williams David R Edgell Carlos T Moraes |
| author_sort | Claudia V Pereira |
| collection | DOAJ |
| description | Abstract Pathogenic mitochondrial DNA (mtDNA) mutations often co‐exist with wild‐type molecules (mtDNA heteroplasmy). Phenotypes manifest when the percentage of mutant mtDNA is high (70–90%). Previously, our laboratory showed that mitochondria‐targeted transcription activator‐like effector nucleases (mitoTALENs) can eliminate mutant mtDNA from heteroplasmic cells. However, mitoTALENs are dimeric and relatively large, making it difficult to package their coding genes into viral vectors, limiting their clinical application. The smaller monomeric GIY‐YIG homing nuclease from T4 phage (I‐TevI) provides a potential alternative. We tested whether molecular hybrids (mitoTev‐TALEs) could specifically bind and cleave mtDNA of patient‐derived cybrids harboring different levels of the m.8344A>G mtDNA point mutation, associated with myoclonic epilepsy with ragged‐red fibers (MERRF). We tested two mitoTev‐TALE designs, one of which robustly shifted the mtDNA ratio toward the wild type. When this mitoTev‐TALE was tested in a clone with high levels of the MERRF mutation (91% mutant), the shift in heteroplasmy resulted in an improvement of oxidative phosphorylation function. mitoTev‐TALE provides an effective architecture for mtDNA editing that could facilitate therapeutic delivery of mtDNA editing enzymes to affected tissues. |
| first_indexed | 2024-03-07T17:37:04Z |
| format | Article |
| id | doaj.art-7a381d0d4b6e4d0eb16577d5be175b52 |
| institution | Directory Open Access Journal |
| issn | 1757-4676 1757-4684 |
| language | English |
| last_indexed | 2024-03-07T17:37:04Z |
| publishDate | 2018-09-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | EMBO Molecular Medicine |
| spelling | doaj.art-7a381d0d4b6e4d0eb16577d5be175b522024-03-02T16:35:03ZengSpringer NatureEMBO Molecular Medicine1757-46761757-46842018-09-01109n/an/a10.15252/emmm.201708084mitoTev‐TALE: a monomeric DNA editing enzyme to reduce mutant mitochondrial DNA levelsClaudia V Pereira0Sandra R Bacman1Tania Arguello2Ugne Zekonyte3Sion L Williams4David R Edgell5Carlos T Moraes6Department of Neurology University of Miami Miller School of Medicine Miami FL USADepartment of Neurology University of Miami Miller School of Medicine Miami FL USADepartment of Neurology University of Miami Miller School of Medicine Miami FL USADepartment of Neurology University of Miami Miller School of Medicine Miami FL USADepartment of Neurology University of Miami Miller School of Medicine Miami FL USADepartment of Biochemistry Schulich School of Medicine and Dentistry University of Western Ontario London ON CanadaDepartment of Neurology University of Miami Miller School of Medicine Miami FL USAAbstract Pathogenic mitochondrial DNA (mtDNA) mutations often co‐exist with wild‐type molecules (mtDNA heteroplasmy). Phenotypes manifest when the percentage of mutant mtDNA is high (70–90%). Previously, our laboratory showed that mitochondria‐targeted transcription activator‐like effector nucleases (mitoTALENs) can eliminate mutant mtDNA from heteroplasmic cells. However, mitoTALENs are dimeric and relatively large, making it difficult to package their coding genes into viral vectors, limiting their clinical application. The smaller monomeric GIY‐YIG homing nuclease from T4 phage (I‐TevI) provides a potential alternative. We tested whether molecular hybrids (mitoTev‐TALEs) could specifically bind and cleave mtDNA of patient‐derived cybrids harboring different levels of the m.8344A>G mtDNA point mutation, associated with myoclonic epilepsy with ragged‐red fibers (MERRF). We tested two mitoTev‐TALE designs, one of which robustly shifted the mtDNA ratio toward the wild type. When this mitoTev‐TALE was tested in a clone with high levels of the MERRF mutation (91% mutant), the shift in heteroplasmy resulted in an improvement of oxidative phosphorylation function. mitoTev‐TALE provides an effective architecture for mtDNA editing that could facilitate therapeutic delivery of mtDNA editing enzymes to affected tissues.https://doi.org/10.15252/emmm.201708084heteroplasmyI‐TevImitochondrial DNAmitoTev‐TALEmonomeric |
| spellingShingle | Claudia V Pereira Sandra R Bacman Tania Arguello Ugne Zekonyte Sion L Williams David R Edgell Carlos T Moraes mitoTev‐TALE: a monomeric DNA editing enzyme to reduce mutant mitochondrial DNA levels EMBO Molecular Medicine heteroplasmy I‐TevI mitochondrial DNA mitoTev‐TALE monomeric |
| title | mitoTev‐TALE: a monomeric DNA editing enzyme to reduce mutant mitochondrial DNA levels |
| title_full | mitoTev‐TALE: a monomeric DNA editing enzyme to reduce mutant mitochondrial DNA levels |
| title_fullStr | mitoTev‐TALE: a monomeric DNA editing enzyme to reduce mutant mitochondrial DNA levels |
| title_full_unstemmed | mitoTev‐TALE: a monomeric DNA editing enzyme to reduce mutant mitochondrial DNA levels |
| title_short | mitoTev‐TALE: a monomeric DNA editing enzyme to reduce mutant mitochondrial DNA levels |
| title_sort | mitotev tale a monomeric dna editing enzyme to reduce mutant mitochondrial dna levels |
| topic | heteroplasmy I‐TevI mitochondrial DNA mitoTev‐TALE monomeric |
| url | https://doi.org/10.15252/emmm.201708084 |
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