Objective: To assess awareness about prolonged sitting and acceptance of interventions to reduce bouts of prolonged sitting. Participants: Undergraduate, graduate, and professional students and faculty from a large public university. Methods: Eight focus groups were conducted with 4 groups of undergraduate students and 4 groups of graduate/professional students. Eight key informant interviews were conducted with faculty. Content analyses were performed using inductive and deductive techniques to elicit common themes. Results: Many students and faculty were not aware of the detrimental effects associated with prolonged sitting. Barriers to movement in university settings included social acceptability, environmental constraints, and academic requirements. Acceptable interventions included: faculty prompts, changes to pedagogy, educational campaigns, and structural changes to classroom designs. Conclusions: University settings represent an opportunity to address issues related to prolonged sitting that may address immediate health implications, as well as establish behaviors and practices that can be continued in work-based settings.