UNLABELLED: Artificial intelligence (AI) is constantly improving the quality of medical procedures. Despite the application of AI in the healthcare industry, there are conflicting opinions among professionals, and limited research on its practical application in Saudi Arabia was conducted. AIM: To assess the nurses' knowledge regarding the application of AI in practice at one of the Ministry of Health hospitals in Saudi Arabia. METHODS: Descriptive cross-sectional research using convenience sampling in January 2023 involving 307 staff nurses, using a single 11-item questionnaire. In addition, 6 closed-ended questions were used to assess the knowledge, possible risks, and advantages of AI. RESULTS: All 307 participants completed the survey and used it for data analysis using SPSS V.25. Kruskal-Wallis and Whitney tests and descriptive statistics were used to identify the significant differences among groups. The study results reveal significant differences between age groups and working locations regarding familiarity with AI and future use of AI. In contrast, a considerable difference exists between licensed years groups regarding familiarity with AI. Surprisingly, education level does not affect AI knowledge. Additionally, the future use of AI is significantly affected by the nurse's gender. LIMITATION: Nurses were not included in previous studies on AI, and most nursing participants need more interest in AI. CONCLUSION: The study's results showed that nurses have positive opinions of AI in the healthcare industry, which will help them speed up procedures and reduce medical errors. AI applications can expand in healthcare by increasing the use of AI in the healthcare industry to improve care quality and encourage academic institutions to develop best practices for deploying AI applications in the healthcare industry.