The convenience and adaptability of CRISPR has led to its rapid and widespread adoption as a toolkit for gene editing, control and labelling. A gap in its exploitation is in the manipulation of organelle genomes, such as in mitochondria. Mutations in mitochondrial DNA (mtDNA) lead to diseases with variable phenotypes and cures are not currently available. In addition, little is known about mtDNA genetics compared to nuclear DNA, mainly because there is a lack of organelle-specific molecular biology tools. Our ambition is to re-engineer CRISPR to allow reliable trafficking to human mitochondria, transport across the double membrane, and reconstitution of activity within the matrix. Our strategy combines protein and RNA design, protein activity analysis, and cell biology.

Project lead: Professor Mark Szczelkun (DNA-protein interactions, CRISPR mechanism, enzymology)

Project team: Professor Ian Collinson (membrane biology, mitochondrial import, structure-function analysis); Dr Jon Lane (mitophagy, autophagy, microscopy); and Dr Marc van der Kamp (protein dynamics, biomolecular simulation)