Work

Protocol to generate genetically engineered organoid-initiated mouse models of esophageal cancer

Year: 2025

Type: article

Abstract: Here, we present a protocol for generating genetically engineered organoid-initiated mouse models of esophageal cancer. We describe steps for isolating mouse esophageal primary cells, organoids culture, construct and virus preparation, and genome editing. We then detail procedures for transplanting engineered organoids into the esophagus, forming squamous cell carcinoma, and tumorigenesis monitoring. The protocol is faster than genetically engineered mouse models. For complete details on the use and execution of this protocol, please refer to Wang et al. 1 • Steps for constructing a primary, orthotopic, esophageal cancer mouse model • Guidance on isolation and organoid development from mouse esophageal epithelial cells • Procedures for organoid passaging, genome editing, and genotyping • Orthotopic transplant of edited organoids into the mouse esophagus by laparotomy Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Here, we present a protocol for generating genetically engineered organoid-initiated mouse models of esophageal cancer. We describe steps for isolating mouse esophageal primary cells, organoids culture, construct and virus preparation, and genome editing. We then detail procedures for transplanting engineered organoids into the esophagus, forming squamous cell carcinoma, and tumorigenesis monitoring. The protocol is faster than genetically engineered mouse models.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.104028