Tools for editing the mammalian mitochondrial genome.

Area of Science:

• Mitochondrial biology
• Gene editing technologies
• Molecular genetics

Background:

• Mitochondrial genome manipulation has been difficult due to limited transformation methods.
• Mitochondrial DNA (mtDNA) mutations can cause disease, often existing as heteroplasmic variants.
• RNA delivery into mitochondria is inefficient, historically limiting CRISPR applications.

Purpose of the Study:

• To review recent advancements in animal mitochondrial DNA (mtDNA) gene editing.
• To highlight novel non-CRISPR techniques for in vivo mtDNA modification.
• To discuss the potential of these tools for research and clinical applications.

Main Methods:

• Utilizing engineered nucleases for targeted double-strand breaks in mtDNA.
• Employing cytidine and adenine deaminases for base editing of mtDNA in vivo.
• Exploring novel, optimized non-CRISPR gene editing strategies.

Main Results:

• Development of effective in vivo gene editing tools for animal mtDNA.
• Demonstration of selective removal or modification of mutant mtDNA variants.
• Emergence of a versatile toolbox for mtDNA research and therapeutic development.

Conclusions:

• Recent breakthroughs enable precise manipulation of animal mtDNA in vivo.
• These gene editing technologies are crucial for studying mitochondrial function.
• The advancements bring potential therapies for mitochondrial disorders closer to reality.