Autosomal dominant polycystic kidney disease (ADPKD) is the most common inherited kidney disease, with a prevalence of 1in 1000 live births. It is characterized by the appearance of cysts in the kidneys and by an increase in the size of the kidneys, leading irrevocably to end-stage renal disease.

Most of the mutations observed in the genes that cause this disease (PKD1 and PKD2) are missense mutations, which makes possible the application of Prime Editing technology to correct these mutations. The correction of the ADPKD-causing mutation should allow a reversion of the ADPKD phenotype, since it has been demonstrated that the kidney is an organ that has plasticity, that is, the correct re-expression of the Pkd1and Pkd2 genes in mouse cystic kidneys results in a rapid reversal of ADPKD.

The overall objective is to use Prime Editing technology to correct the ADPKD-causing mutation in two different mouse models. The specific objectives are: 1) In silico design of pegRNAs; 2) In vitro testing of pegRNAs in mouse embryonic fibroblast (MEFs); 3) Bioinformatics study and selection of the pegRNAs with the highest editing efficiency; 4) In vivo editing of the two mouse models with the selected pegRNAs.

After in silico design of pegRNAs and isolation of MEFs, several in vitro nucleofection experiments with different amounts of pegRNAs and Prime Editing plasmids have been performed in these cells. So far, the editing efficiency achieved is low.

New transfection experiments using mRNA instead of plasmid are being carried out to increase the editing efficiency.