A scalable embryonic stem cell-based platform for efficient generation of mitochondrial DNA mutant mice.
Saved in:
| Title: | A scalable embryonic stem cell-based platform for efficient generation of mitochondrial DNA mutant mice. |
|---|---|
| Authors: | Fan, Weiwei1 wfan@salk.edu, Oh, Tae Gyu1,2, Crossley, Lillian1, Robbins, Hunter1, He, Mingxiao1, Dai, Yang1, Truitt, Morgan L.1, Atkins, Annette R.1, Downes, Michael1, Evans, Ronald M.1 evans@salk.edu |
| Source: | Proceedings of the National Academy of Sciences of the United States of America. 4/14/2026, Vol. 123 Issue 15, p1-8. 17p. |
| Subjects: | Mitochondrial DNA, Embryonic stem cells, Mitochondrial DNA abnormalities, Reactive oxygen species, Laboratory mice, Mitochondria, Germ cells, Oxidative phosphorylation |
| Abstract: | Mitochondria are central to energy metabolism and cellular signaling, and mutations in mitochondrial DNA (mtDNA) can disrupt these processes and contribute to human disease. However, progress in defining how mtDNA variation influences adaptation, pathophysiology, and disease susceptibility has been limited by the lack of suitable animal models. Although recent base-editing approaches enable direct mtDNA modification, their low efficiency restricts the generation of diverse models reflecting human mtDNA variation. Here, we develop a scalable embryonic stem (ES) cell-based platform for efficient production of mtDNA mutant mice. Random mutagenesis using an error-prone mtDNA polymerase generates a broad spectrum of mtDNA mutations, which are transferred into ES cells via a multiplexed cybrid fusion strategy coupled with sensitive mutation detection. Optimized ES cell-embryo aggregation enables robust contribution of mtDNA mutant ES cells to host embryos, producing chimeric mice with germline transmission. Using this platform, we generate a library of 155 donor fibroblast lines carrying distinct homoplasmic single-nucleotide mtDNA mutations that produce diverse mitochondrial phenotypes, including impaired oxidative phosphorylation, increased reactive oxygen species, and altered mitochondrial membrane potential. We further generate 34 female C57BL/6 ES cell lines harboring 18 mtDNA mutations across a range of heteroplasmy levels, yielding multiple chimeric mice and achieving germline transmission for one mutation. These data reveal a strong correlation between mitochondrial function and early embryonic development, suggesting a minimal energetic threshold required for normal development. This scalable resource enables systematic investigation of mtDNA variation in physiology, adaptation, disease mechanisms, and therapeutic development. [ABSTRACT FROM AUTHOR] |
| Copyright of Proceedings of the National Academy of Sciences of the United States of America is the property of National Academy of Sciences and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) | |
| Database: | Engineering Source |
| FullText | Text: Availability: 0 |
|---|---|
| Header | DbId: egs DbLabel: Engineering Source An: 193036263 AccessLevel: 6 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 0 |
| IllustrationInfo | |
| Items | – Name: Title Label: Title Group: Ti Data: A scalable embryonic stem cell-based platform for efficient generation of mitochondrial DNA mutant mice. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Fan%2C+Weiwei%22">Fan, Weiwei</searchLink><relatesTo>1</relatesTo><i> wfan@salk.edu</i><br /><searchLink fieldCode="AR" term="%22Oh%2C+Tae+Gyu%22">Oh, Tae Gyu</searchLink><relatesTo>1,2</relatesTo><br /><searchLink fieldCode="AR" term="%22Crossley%2C+Lillian%22">Crossley, Lillian</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Robbins%2C+Hunter%22">Robbins, Hunter</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22He%2C+Mingxiao%22">He, Mingxiao</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Dai%2C+Yang%22">Dai, Yang</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Truitt%2C+Morgan+L%2E%22">Truitt, Morgan L.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Atkins%2C+Annette+R%2E%22">Atkins, Annette R.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Downes%2C+Michael%22">Downes, Michael</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Evans%2C+Ronald+M%2E%22">Evans, Ronald M.</searchLink><relatesTo>1</relatesTo><i> evans@salk.edu</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22Proceedings+of+the+National+Academy+of+Sciences+of+the+United+States+of+America%22">Proceedings of the National Academy of Sciences of the United States of America</searchLink>. 4/14/2026, Vol. 123 Issue 15, p1-8. 17p. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22Mitochondrial+DNA%22">Mitochondrial DNA</searchLink><br /><searchLink fieldCode="DE" term="%22Embryonic+stem+cells%22">Embryonic stem cells</searchLink><br /><searchLink fieldCode="DE" term="%22Mitochondrial+DNA+abnormalities%22">Mitochondrial DNA abnormalities</searchLink><br /><searchLink fieldCode="DE" term="%22Reactive+oxygen+species%22">Reactive oxygen species</searchLink><br /><searchLink fieldCode="DE" term="%22Laboratory+mice%22">Laboratory mice</searchLink><br /><searchLink fieldCode="DE" term="%22Mitochondria%22">Mitochondria</searchLink><br /><searchLink fieldCode="DE" term="%22Germ+cells%22">Germ cells</searchLink><br /><searchLink fieldCode="DE" term="%22Oxidative+phosphorylation%22">Oxidative phosphorylation</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: Mitochondria are central to energy metabolism and cellular signaling, and mutations in mitochondrial DNA (mtDNA) can disrupt these processes and contribute to human disease. However, progress in defining how mtDNA variation influences adaptation, pathophysiology, and disease susceptibility has been limited by the lack of suitable animal models. Although recent base-editing approaches enable direct mtDNA modification, their low efficiency restricts the generation of diverse models reflecting human mtDNA variation. Here, we develop a scalable embryonic stem (ES) cell-based platform for efficient production of mtDNA mutant mice. Random mutagenesis using an error-prone mtDNA polymerase generates a broad spectrum of mtDNA mutations, which are transferred into ES cells via a multiplexed cybrid fusion strategy coupled with sensitive mutation detection. Optimized ES cell-embryo aggregation enables robust contribution of mtDNA mutant ES cells to host embryos, producing chimeric mice with germline transmission. Using this platform, we generate a library of 155 donor fibroblast lines carrying distinct homoplasmic single-nucleotide mtDNA mutations that produce diverse mitochondrial phenotypes, including impaired oxidative phosphorylation, increased reactive oxygen species, and altered mitochondrial membrane potential. We further generate 34 female C57BL/6 ES cell lines harboring 18 mtDNA mutations across a range of heteroplasmy levels, yielding multiple chimeric mice and achieving germline transmission for one mutation. These data reveal a strong correlation between mitochondrial function and early embryonic development, suggesting a minimal energetic threshold required for normal development. This scalable resource enables systematic investigation of mtDNA variation in physiology, adaptation, disease mechanisms, and therapeutic development. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of Proceedings of the National Academy of Sciences of the United States of America is the property of National Academy of Sciences and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
| PLink | https://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=egs&AN=193036263 |
| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1073/pnas.2535453123 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 17 StartPage: 1 Subjects: – SubjectFull: Mitochondrial DNA Type: general – SubjectFull: Embryonic stem cells Type: general – SubjectFull: Mitochondrial DNA abnormalities Type: general – SubjectFull: Reactive oxygen species Type: general – SubjectFull: Laboratory mice Type: general – SubjectFull: Mitochondria Type: general – SubjectFull: Germ cells Type: general – SubjectFull: Oxidative phosphorylation Type: general Titles: – TitleFull: A scalable embryonic stem cell-based platform for efficient generation of mitochondrial DNA mutant mice. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Fan, Weiwei – PersonEntity: Name: NameFull: Oh, Tae Gyu – PersonEntity: Name: NameFull: Crossley, Lillian – PersonEntity: Name: NameFull: Robbins, Hunter – PersonEntity: Name: NameFull: He, Mingxiao – PersonEntity: Name: NameFull: Dai, Yang – PersonEntity: Name: NameFull: Truitt, Morgan L. – PersonEntity: Name: NameFull: Atkins, Annette R. – PersonEntity: Name: NameFull: Downes, Michael – PersonEntity: Name: NameFull: Evans, Ronald M. IsPartOfRelationships: – BibEntity: Dates: – D: 14 M: 04 Text: 4/14/2026 Type: published Y: 2026 Identifiers: – Type: issn-print Value: 00278424 Numbering: – Type: volume Value: 123 – Type: issue Value: 15 Titles: – TitleFull: Proceedings of the National Academy of Sciences of the United States of America Type: main |
| ResultId | 1 |
