Yeasts as a cell model for studying Huntinghton's diseaseErzar, Iza (Avtor) Skočaj, Matej (Mentor) Xin, Chen (Komentor) Huntington's diesasehuntingtinSaccharomyces cerevisiaeyeastmodelHuntington's disease (HD) is a neurodegenerative disorder (ND) caused by mutation in huntingtin (HTT) gene. To date there has been no approved cure for HD, which is mainly due to the fact that the molecular mechanisms of the disease have not been well studied and defined. Therefore, stable and reliable model organisms are important for disease studies. Yeast is potential model organism for studying this disease, since both in yeast and in the normal course of the disease, mutated HTT misfolds and forms aggregates within the cell. In this study, yeast cell models stably and constitutively expressing normal and mutated huntingtin (HTT) protein have been constructed. Since yeast has no homolog of huntingtin CRISPR Cas9 system has been used for gene integration. The procedures for phenotypic verification of the models are described, namely growth rate and viability, which includes the determination of CFU (colony-forming-units), spotting assay, and measurement of dead cells using propidium iodide (PI) and flow cytometer. As the GFP (green-fluorescence-protein) is fused at the N-terminal region of HTT protein, its expression and the formation of characteristic aggregates of the mutated protein are observed under the fluorescence microscope. Troubleshooting is also included since difficulties can (and did occur) occur during model construction. The obtained models have great potential as a platform for understanding the disease mechanisms and further enhance their potential for drug screening. Additional research will be needed for the phenotypical characteristics and other properties of these models.20222022-08-29 10:04:24Diplomsko delo/naloga139038VisID: 208157COBISS_ID: 119779331sl