Abstract: The cell-free protein synthesis (CFPS) system has received widespread attention as a general tool for basic research, applied science, and product development. Significant advancements have been achieved in synthesizing target proteins using CFPS systems in the fields of food, medicine, and synthetic biology. Among the existing CFPS systems, the wheat germ-based cell-free protein synthesis (WG-CFPS) system stands out due to its wide source of raw materials, high protein yield, and suitability for expressing complex eukaryotic proteins and membrane proteins. It has been identified as a cost-effective and promising candidate for commercial development. This review summarizes the current research status related to the WG-CFPS system and discusses various optimization strategies and their outcomes within this system. In addition, the current application of WG-CFPS system in food, medicine and other fields is summarized and sorted out, and its future challenges and development are prospected.