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Updated: Mar 29, 2026
An In Vitro Approach to Study Mitochondrial Dysfunction: A Cybrid Model
Published on: March 9, 2022
One-step generation of heritable mitochondrial DNA multiplex-engineered rats using DddA-derived cytosine base editor
1State Key Laboratory of Respiratory Health and Multimorbidity, NHC Key Laboratory of Human Disease Comparative Medicine, Key Laboratory of Pathogen Infection Prevention and Control Ministry of Education, Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
Researchers created the first heritable multi-site mitochondrial DNA (mtDNA) mutant rats using TALE-based genome editing. This breakthrough provides a vital model for studying mitochondrial diseases and developing new therapies.
Area of Science:
- Genetics and Genomics
- Molecular Biology
- Disease Modeling
Background:
- Mitochondrial DNA (mtDNA) mutations are linked to numerous diseases.
- Studying diseases with multiple mtDNA mutations is challenging due to the lack of appropriate animal models.
- Matrilineal inheritance of mtDNA prevents traditional breeding for creating multi-mutation models.
Purpose of the Study:
- To develop a novel animal model for studying mitochondrial diseases.
- To establish rats harboring multiple mitochondrial DNA mutations.
- To investigate the efficacy and safety of TALE-based mitochondrial genome editing.
Main Methods:
- Utilized TALE-based mitochondrial genome editing tools (DdCBE).
- Microinjected mixed DdCBE plasmids into one-cell-stage zygotes to generate mutant rats.
- Assessed mtDNA editing efficiency and off-target effects using deep and long-read sequencing.
Main Results:
- Successfully generated double- and triple-site mtDNA mutant rats with editing efficiencies up to 58.5%.
- Confirmed stable germline transmission of the induced mtDNA mutations.
- Demonstrated minimal off-target effects in both mitochondrial and nuclear genomes in vivo.
Conclusions:
- This study reports the first successful generation of heritable multi-site mtDNA mutant rats.
- These novel mutant rats serve as a valuable preclinical model for mitochondrial disorders.
- The findings pave the way for elucidating disease mechanisms and developing targeted therapeutics.
