Hi !
I'd like to induce a DNA repair mechanism by homology with a donor sequence on mitochondrial DNA. Have you already succeeded in sending a template sequence to the mitochondria? If so, by what means (lipofectamine transfection, RNAi max, etc.)?
Thank you...
You can add a mitochondrial membrane localization sequence. See https://www.embopress.org/doi/epdf/10.1038/embor.2013.168?src=getftr. They used the aa101-152 peptide of FIS1 protein for mitochondrial targeting. But the peptide sequence only works post-translationally and I'm not sure if the peptide sequence is compatible with DNA delivery, maybe with a cleavable tag.
There are also mitochondrial dyes in the literature space for possible chemical modification that can assist mitochondrial localization. Modification chemically / through DNA might combine well with lipofectamine to deliver DNA directly to the mitochondria.
This is one of the most challenging problems of genetics. It's challenging to alter the mitochondrial genome as it lacks the machinery for homologous recombination. You can easily target a protein into the mitochondria but not insert any RNA or DNA into the mitochondria. Even CRISPR needs some guide RNA to manipulate the mtDNA. Please see this link
https://www.nature.com/articles/s43586-023-00200-7
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