There is a strong clinical need for novel therapeutic options for pancreatic ductal adenocarcinoma (PDAC), where the 5-year overall survival remains poor at 15% with limited treatment options beyond chemotherapy (1, 2). Increased expression of PRMT5 in PDAC is associated with a worse prognosis and its inhibition in PDAC preclinical models have shown to be an effective therapy (3, 4). However, it remains unclear how the diverse number of cellular processes PRMT5 inhibition affects contribute to its role in cancer growth inhibition (5). Interestingly, the more alternative splicing PRMT5 inhibition induces, the greater the effect on tumour growth inhibition (6, 7). Thus, this project aims to uncover the mechanism of action of PRMT5 inhibition in PDAC and in doing so advance the development of this novel therapeutic class (targeting alternative splicing) for this disease. To achieve this aim, we have performed whole transcriptome sequencing to determine time dependent PRMT5 inhibitor-induced differential mRNA expression and changes in pre-mRNA alternative splicing in two cell line models. These cell line models represent Moffitt’s basal (MiaPaca2) and classical (AsPC1) molecular subtype (8) where emerging evidences suggest there are subtype-specific responses. In addition, a CRISPR screen was performed to assess genes necessary to respond to PRMT5 inhibition and potential mechanisms leading to synergistic growth inhibition. Updated analysis results combining both unbiased approaches will be presented which identify a list of relevant genes for functional validation towards uncovering PRMT5 inhibitor’s mechanism of action.