siRNA-Mediated TDP-43 Nuclear Depletion in NSC-34 Cells to generate ALS-like phenotype for pharmacological intervention

This thesis investigates TDP43 nuclear depletion in motor neuron-like cells with the aim of generating ALS-like phenotype in-vitro model for the evaluation of possible pharmaceutical intervention to restore cell vitality and cell differentiation capability. 97% of ALS cases exhibit TDP43 proteinopathy leading to misfolded aggregates with prion-like behavior, although the specific contribution of nuclear deficiency of TDP43 in the ALS phenotype are not yet fully understood, it might be targeted to ameliorate pathological phenotype. To generate this model, siRNA-mediated transfection was performed on NSC34 cells (neuroblastoma x spinal cord hybrid line). The experimental workflow included quantification of protein silencing via BCA, SDS-PAGE, Western blot and qualitative assays through immunocytochemistry paired with confocal fluorescent microscopy. Preliminary results showed a 22% average expression reduction, prompting further investigation to raise the silencing expression and evaluate cell vitality for pharmaceutical treatment.