Background: Gut microbiome is a vast microorganism's ecosystem that resides in the human gastrointestinal tract. Microbiota can have a complex crosstalk with the central nervous system, creating "gut-brain axis", playing a crucial role in neuropsychiatric disorders pathogenesis. Aim: This study aimed to assess knowledge about human gut microbiota, dysbiosis and gut-brain interplay in neuropsychiatric disorders among Saudi medical students.Methods: This is a descriptive cross-sectional study using questionnaires to collect the data, targeting senior medical students and interns in Saudi Arabia from November 2022 to January 2023. Results: Overall, our study shows that our students possess adequate general knowledge about microbiota, dysbiosis and probiotics; however, they lack sufficient knowledge regarding gut-brain axis and dysbiosis role in the context of neuropsychiatric diseases pathogenesis and management. Conclusion: Developing the medical curricula and adopting more modalities of learning strategies to discuss the most recent scientific advances in different medical topics are crucial to enhance medical students' knowledge acquisition, thus providing adequate recommendation to their future patients and community.