Nanoformulations of prime editing ribonucleoproteins

The development of the CRISPR/Cas9 system as a molecular tool for targeted DNA modification has revolutionized biomedical research and sparked great hopes for novel therapies. The functional unit of CRISPR/Cas9 consists of ribonucleoproteins (RNPs), which act as controllable "gene scissors" to cut DNA at desired sequences. In recent years, advancements in the conventional, naturally occurring CRISPR/Cas9 system have enhanced its precision and versatility, adapting it to specific medical requirements. One such innovation is "Prime Editing" (PE), which enables a range of genetic modifications, including short deletions, insertions, and nucleobase conversions, while simultaneously reducing the risk of off-target errors. It is estimated that up to 89% of known disease-associated genetic variants could theoretically be corrected using PE. However, for the treatment of human diseases, the components must be safely and efficiently delivered into the nucleus of desired target cells, which currently represents a major hurdle for widespread application. The project "Nanoformulations of Prime Editing Ribonucleoproteins" addresses this challenge by developing nanoscale carrier systems for PE RNPs and systematically investigating them in cell models. These carriers are based on material classes that have already demonstrated excellent suitability for the cellular transport of other biomolecules and are being specifically optimized for PE RNPs: ionizable lipo-oligomers and lipid nanoparticles (LNPs). Using a combination of complementary methods, the nanoformulations will be characterized in terms of their efficiency in delivering PE RNPs into cells, mediating desired DNA modifications, and minimizing unwanted effects. In this way, relevant cellular transport processes can be deciphered, and optimal carrier properties for future PE therapies can be identified.