OBJECTIVES: A cricothyrotomy represents an emergency procedure that may be considered a last option for securing the airway. While fortunately rare, it is important to note that such invasive procedures must be mastered if they are to be used. Therefore, regular training is essential to gain routine. The aim of the present study was to investigate whether professional groups with different levels of experience with the procedure succeed in learning the procedure with a virtual reality trainer. MATERIALS AND METHODS: In a multicenter approach, 146 employees with four different professional backgrounds-otorhinolaryngologists, anesthesiologists, emergency physicians and certified nurses-were included in the study. The participants were required to complete a virtual reality (VR) cricothyrotomy scenario in three consecutive runs, and the time required and errors in the procedure were recorded. The training experience was subsequently evaluated subjectively using a questionnaire. RESULTS: The study included 146 participants with an average age of 33 years and an average of 5 years of professional experience. The majority of participants (74%) reported an improvement in the speed of the procedure and in the procedural steps (87%). These subjective improvements were confirmed objectively by the time required for completion of the procedure and the points achieved. Gaming experience had a significant effect on both the score (p = 0.023) and procedure time (p = 0.039), whereas age and medical specialization did not. Real-life experience with cricothyrotomy had no significant effect on performance in VR. CONCLUSION: Virtual reality provides an effective method for training healthcare professionals in cricothyrotomy, regardless of their specialty or prior experience. The participants showed significant improvements in both the speed and accuracy of the procedure after training, regardless of their prior experience or medical background. Further research is necessary to assess the benefits of VR simulation for training cricothyrotomy in real-world procedures. TRIAL REGISTRATION: DRKS00031736, registered on the 20th of April 2023.