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

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.