Anvi Murarka

Validating Genetic Modifiers of TAU Clearance Using an Arrayed Screening Approach

Faculty PI: Kenneth Kosik
Alzheimer's disease and related neurodegenerative disorders are characterized by the buildup of the tau protein, which misfolds and clumps together in neurons, disrupting their normal function. The brain has natural ways of clearing this damaged tau before it accumulates, and promoting this clearance process could be a promising therapeutic strategy. However, the specific genes that control tau clearance and the mechanism by which tau spreads are not yet fully understood. To address this gap, a pilot CRISPR screen tested candidate genes to see whether removing them affected the cell's ability to break down tau. This screen identified several genes, including NPC2, RAB7A, MTOR, SQSTM1, PSMB5, and USP14, each involved in a different cellular degradation system. To build on and validate these findings, we are conducting individual gene knockdowns to see if removing each gene causes tau fibrils to build up, confirming the gene’s role in tau clearance. Candidate genes were first silenced at varying doses in a neuroglioma cell line, and the concentration producing the strongest gene silencing was identified for each. These optimized conditions will next be used to directly visualize whether tau fails to clear, and instead accumulates, following gene knockdown. In parallel, we are extending this work into a more physiologically relevant model by developing a stem cell-derived neural cell line that allows fluorescent tau production to be switched on and off on demand. Together, this work lays the groundwork for identifying the specific genes responsible for tau clearance and potential targets for future Alzheimer's therapies.