Développement d'un ribozyme spécifique à l'ARNm de Tau pour contrer les Tauopathies

Authors: Eyoum Jong, Laura
Advisor: Planel, EmmanuelLévesque, Georges
Abstract: One of the main causes of Alzheimer’s disease and Tauopathies is the presence of neurofibrillary tangles (NFT). NFT consist in intracellular aggregation of the abnormally hyperphophorylated protein Tau. Several studies have shown that NFT are associated with the pathogenesis of neurodegenerative disorders and neurotoxicity. Moreover, it has been demonstrated that decreasing the level of Tau protein could prevent cognitive deficits in mouse models. Based on these studies, our hypothesis is that reducing the level of total Tau in the brain could decrease NFTs and delay the pathology. Our objective is to design a molecule that will target directly Tau mRNA instead of the hyperphosphorylated protein. We developed a modified delta-ribozyme, the SOFA (Specific On/Off Adaptor) ribozyme, which is able to cleave the Tau mRNA. Our ribozyme is composed of three components: the blocker, the biosensor, and the effector. We have designed delta-ribozymes that target constitutive exons of Tau mRNA as well as the exon 10 specific of Tau 4R. Therefore, we can target all the Tau isoforms. In this project, we first synthesized the delta-ribozyme by molecular cloning and we confirmed its effect by in vitro cleavage. Secondly, by transfecting it in neuronal cells we characterized its effects on Tau mRNA by RT-PCR. Finally, we produced AAV viruses and infected neuronal cells and characterized its effects on Tau mRNA by RT-PCR. This therapeutic approach is based on specificity and efficiency of the SOFA-ribozymes, which identify and cut the Tau mRNA. Thus, in this project we identified ribozymes 350 and 395 as potential good candidates able to cleave the Tau mRNA.
Document Type: Mémoire de maîtrise
Issue Date: 2018
Open Access Date: 18 April 2019
Permalink: http://hdl.handle.net/20.500.11794/34496
Grantor: Université Laval
Collection:Thèses et mémoires

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