BACKGROUND: AI can improve medical practice, address staff shortages, and enhance diagnostic efficiency. The ChatGPT of Open AI, launched in 2022, uses AI in medical education. However, the long-term impact is uncertain, and integration varies globally, particularly in the Middle East. AIM: To explore the knowledge, practices, and attitudes of nursing students in Palestinian universities regarding AI, specifically the use of ChatGPT. METHODOLOGY: A cross-sectional design was used to conduct this study. The study was performed at 8 private and governmental universities in the West Bank, Palestine, from 1st May 2024 to 30 May 2024, and 304 nursing students participated. RESULTS: The study revealed that 84.5% of nursing students at Palestinian universities were aware of AI technology, yet 69.9% lacked formal education or training related to ChatGPT. Despite this gap, 79% supported the integration of AI into nursing curricula and specialized training programs, reflecting strong optimism about its role in education and healthcare. While 58.6% had used AI in their coursework and 68.1% felt comfortable with technology, disparities in proficiency and access remain key barriers to effective AI integration. Major challenges to AI adoption in Palestine include insufficient training, the absence of AI-focused curricula, and financial constraints, underscoring the need for institutional and pedagogical reforms. Concerns about AI's reliability, costs, and potential diagnostic errors persist, emphasizing the complexities of its integration into nursing education and practice. CONCLUSION: This study highlights the knowledge, attitudes, and practices of Palestinian nursing students regarding AI and ChatGPT. It reveals that, despite growing awareness, the lack of formal education on AI underscores the need for comprehensive curricula. While students' express optimism about AI's potential in healthcare, concerns about its reliability and integration persist. The study also reveals that barriers such as inadequate training, limited curricula, and financial constraints must be addressed to effectively integrate AI into nursing education and prepare students for its expanding role in healthcare.