RAREHOPE

Friedreich Ataxia, which is one of the common ataxia varieties worldwide, is a neurodegenerative disease that occurs when GAA.TTC repeats occur more than necessary in the 1st intron region, which is common from mutations in the FXN gene. Increased GAA codon repeat causes DNA methylation and consequently gene silencing. The main consequence of GAA codon repeat is reduced production of frataxin protein, which has mitochondrial function. GAA codon repeat occurs in the GAA repeat instability region. In this project, it is aimed to cut the GAA repeat instability region using the CRISPR-Cas9 knockout system. The production of frataxin protein was aimed by giving donor DNA which has a 12-repeat GAA codon found in normal individuals to the cut area. For this section, 2 plasmid systems and 2 transport systems based on these plasmids were designed. Two plasmid systems containing 2 sgRNA and 1 SpCas9 CRISPR elements were designed to cut the GAA repeat site, and creation of a ribonucleoprotein was considered for the first project. While considering the use of non-viral cationic lipids to deliver one of these systems to target cells; Virus-mediated delivery, one of the viral delivery systems, was selected for delivery of the second plasmid, and the use of adenovirus was planned. It is aimed to use the FKBP-FRB protein pair with the presence of rapamycin antibiotic in order to connect the cut regions correctly in the first project. As a result of the study, it is aimed to prevent the symptoms of Friedreich’s Ataxia disease by ceasing long GAA repeats.