Tauopathies are neurodegenerative diseases associated with abnormal Tau protein accumulation, which leads to neuronal dysfunction and death, and dementia. To date, over 60 mutations in the gene coding Tau (MAPT) have been linked to pathological accumulation, microtubule instability, organelle trafficking defects, and synaptic dysfunction. A key feature of tauopathies is “selective neuronal vulnerability,” meaning that abnormal Tau differentially affects specific neuronal subtypes. Currently, there are no effective therapies to halt disease progression. Thus, we aimed to design a tool to selectively reduce Tau in affected neurons and assess its therapeutic potential. Here, we evaluated the phenotypes associated with the V337M Tau mutation in patient-derived iPSCs glutamatergic neurons. Using live-cell fluorescence imaging, we assessed lysosomal axonal transport and observed altered distributions of retrograde and anterograde segmental velocities in V337M neurons compared with wild-type. Preliminary calcium imaging experiments further revealed reduced ΔF values and lower signal frequency in V337M neurons, suggesting electrophysiological impairment. In addition, we developed lentiviral vectors carrying microRNAs under the CAMKII promoter to selectively reduce Tau in glutamatergic neurons and validated their efficacy in wild-type human neurons. Future studies will evaluate whether this approach can reverse disease-associated phenotypes in V337M neurons and other Tau mutants.