Abstract
Efficient generation and correction of mutations in mitochondrial DNA (mtDNA) is challenging. Here, through embryonic injection of an mtDNA adenine base editor (eTd-mtABE), Leigh syndrome rat models were generated efficiently (up to 74%) in the F0 generation, exhibiting severe defects. To correct this mutation, a precise mtDNA C-to-T base editor was engineered and injected into mutated embryos. It achieved restoration of wild-type alleles to an average of 53%, leading to amelioration of disease symptoms.
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Data availability
HTS data were deposited to the National Center for Biotechnology Information Sequence Read Archive database under accession code PRJNA1252023. There are no restrictions on data availability. Source data are provided with this paper.
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Acknowledgements
We thank Y. Zhang from the Flow Cytometry Core Facility of School of Life Sciences at ECNU and support from the ECNU Public Platform for innovation (011). We thank L. Ji (HAVAS) for designing schematic diagrams. This work was partially supported by grants from the National Key R&D Program of China (2023YFC3403400 and 2023YFE0209200 to D.L.; 2024YFC3407900 to L.C.), National Natural Science Foundation of China (32025023, 32230064 and 32311530111 to D.L.; 31930016 to W.W.; 82230002 to M.L.), Innovation Program of Shanghai Municipal Education Commission (2019-01-07-00-05-E00054 to D.L.), Shanghai Municipal Commission for Science and Technology (24J22800400 to D.L.), Young Elite Scientist Sponsorship Program by China Association for Science and Technology (2023QNRC001 to L.C.), Shanghai Oriental Talent Plan (QNZH2024131 to L.C.), Fellowship of China Postdoctoral Science Foundation (8206400139 to Z.Y.) and Lingang Laboratory. D.L. is a Shanghai Academy of Natural Sciences Exploration Scholar.
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L.C. and D.L. designed the experiments. L.C., C.L., M.H., M.Y., H. Huang, D.G., X.G., Y.L., L.Y., L.G. and H. Han performed the experiments. L.C., C.L., M.H., M.Y., H. Huang, D.G., X.G., Z.C., Z.Y., W.W., M.L. and D.L. analyzed the data. L.C. and D.L. wrote the paper with input from all authors. L.C. and D.L. supervised the research.
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The authors have submitted patent applications based on the results reported in this study (L.C., D.L., M.H. and C.L.). The other authors declare no competing interests.
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Supplementary Figs. 1–5 and Note.
Supplementary Tables 1–3
Primer sequences used in this study, target sites used in this study and general architecture and DNA or amino acid sequences of plasmids used in this study.
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Source data for Supplementary Figs. 1–5.
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Chen, L., Luan, C., Hong, M. et al. A mitochondrial disease model is generated and corrected using engineered base editors in rat zygotes. Nat Biotechnol (2025). https://doi.org/10.1038/s41587-025-02684-y
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DOI: https://doi.org/10.1038/s41587-025-02684-y
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