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Previous studies have shown that three factors influence fresh-cut produce safety from farm to fork: (1) post-harvest practices in processing facilities, (2) employees’ handling practices in retail facilities, and (3) consumers’ handling practices in domestic kitchens or cooking facilities. However, few studies have examined consumers’ food safety knowledge, risk perceptions, and their handling practices associated with fresh-cut produce. To fill this gap, the present study conducted a nationwide survey to assess U.S. consumers’ food safety knowledge, practices, and risk perception associated with fresh-cut produce among various demographic groups and investigated factors influencing consumers’ food safety practices related to fresh-cut produce. The results showed that consumers lack the knowledge and safe handling practices toward fresh-cut produce regarding storage hierarchy, surface cleaning and sanitizing, and time and temperature control of fresh-cut produce. The men and millennial consumers exhibit a lower level of safe fresh-cut produce handling practices. In addition, a significant interaction was observed between food safety knowledge and risk perceptions on consumers’ fresh-cut produce handling practices, such that food safety knowledge can transfer to practice more effectively for consumers with high levels of risk perception. The results can be utilized to design effective consumer food safety education tools for targeted audiences.