Alterations in mitochondrial base editors enhance targeted editing efficiency for mouse model generation
Mitochondrial DNA (mtDNA) base editors are powerful tools for investigating mitochondrial diseases. However, their editing efficiency can vary significantly depending on the target site within the mtDNA. In this study, we developed two improved versions of the mitochondrial adenine base editor (Hifi-sTALED and αnHifi-sTALED) by modifying components other than the TadA8e-V28R deaminase variant. These enhancements significantly increased editing efficiency while preserving minimal off-target effects across the transcriptome. Using these optimized editors, we achieved improved mtDNA editing in mouse embryos and successfully generated <i>mt-Rnr1</i> mutant mice with high heteroplasmic loads. Functional analyses revealed that the <i>mt-Rnr1</i> mutation impaired mitochondrial function, as indicated by reduced ATP production and decreased oxygen consumption rate (OCR). These findings demonstrate the utility of the enhanced base editors in generating mitochondrial disease models and advancing research in mitochondrial genetics.
Authors
- Jiyoung Bae (ORCID: https://orcid.org/0000-0001-7872-7689)
- Soo-Ji Park (ORCID: https://orcid.org/0009-0003-0417-7487)
- Sol Pin Kim (ORCID: https://orcid.org/0000-0003-1625-5756)
- Yun Kyu Oh (ORCID: https://orcid.org/0000-0001-8632-5743)
- Chang Geun Choi
- Je Kyung Seong (ORCID: https://orcid.org/0000-0003-1177-6958)
- Hyejung Cha (ORCID: https://orcid.org/0000-0003-0140-7067)
- Lark Kyun Kim (ORCID: https://orcid.org/0000-0001-5983-4470)
- Kyoungmi Kim (ORCID: https://orcid.org/0000-0003-0941-806X)
- Hyunji Lee (ORCID: https://orcid.org/0000-0001-9957-9275)
- Do Hyun Kim (ORCID: https://orcid.org/0000-0002-9248-5572)
- Jieun Kim (ORCID: https://orcid.org/0000-0002-6498-5801)
- Jong Cheol Park (ORCID: https://orcid.org/0000-0002-8859-5111)
- Sanghun Kim (ORCID: https://orcid.org/0000-0003-2745-1728)
- Seongho Hong (ORCID: https://orcid.org/0000-0002-8787-6974)
- Sungmo Jung
- Seung Hee Choi
- Soo Kyung Kang
- Su Bin Lee
Institutions
- Seoul National University (KR)
- Cancer Research Institute (US)
- Yonsei University (KR)
- Korea University (KR)
- Institute of Molecular Biology (AM)
- Gangnam Severance Hospital (KR)
- Korea University (JP)
Publication Details
- Journal
- Molecular Therapy — Nucleic Acids
- Published
- 2025-08-11
- DOI
- https://doi.org/10.1016/j.omtn.2025.102678
- Primary Topic
- Mitochondrial Function and Pathology
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Korea Institute for Advancement of Technology
- National Research Foundation of Korea
- Ministry of Science and ICT, South Korea