Manipulation of Gene Expression in Mouse Pancreas via Intraductal Delivery of Adeno-Associated Viral Vectors

The rising global incidence of pancreatitis, pancreatic cancer, and diabetes has increased the need for efficient in vivo gene manipulation approaches to study the pancreas and develop new therapies. Although transgenic mouse models are widely used, they are time-consuming and costly to generate and maintain. Systemic viral delivery methods offer greater flexibility but often lack pancreatic specificity and require high viral doses. Here, we describe a streamlined protocol for intrapancreatic ductal delivery of adeno-associated viruses (AAVs) for targeted gene delivery. Our protocol requires standard surgical equipment and can be implemented in most laboratories. Specifically, we adopted a clamping strategy at the hepatopancreatic duct near the liver, as well as beneath the major duodenal papilla at the duodenum. This strategy exposes the duodenal papilla, facilitating viral delivery, preventing backflow, and enabling efficient pancreatic transduction at lower viral doses. Overall, this method provides a fast, simple, and effective approach for pancreas-targeted gene manipulation, facilitating preclinical studies of pancreatic biology and disease. Key features • Rapid, pancreas-specific in vivo gene manipulation using simple rodent surgical techniques. • Efficient gene manipulation can be achieved with lower viral doses while minimizing off-target effects. • AAVs trigger minimal adverse complications, and the surgery is well-tolerated in mice. • This method can be combined with traditional genetic manipulation and lineage tracing to enhance studies of gene function or pancreatic diseases.