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It is an ethical imperative that faculty and staff develop a level of cultural competence in order to work effectively with increasingly diverse student bodies, as well as to prepare students to effectively participate in a diverse, global workforce. Online learning offers a promising way to provide meaningful, in-depth diversity and inclusion education to faculty and staff who typically have limited time to devote to professional development activities. However, little is known about the effectiveness of such learning interventions. The current project utilized a pretest-posttest design to examine the effectiveness of a 4-week instructor-led web-based course facilitated on Blackboard Learning Management System. Effectiveness was defined as increasing participants’ cultural competence in terms of cognitive, affective, and behavioral learning related to building a more inclusive campus environment. The findings of this study provide initial evidence for the use of this online course to support the development of faculty and staff cultural competence. Results indicate that participants developed a greater understanding of the value of diversity, an increased openness to learning about other cultures, and a greater awareness of their social privileges. Results also suggest that individuals’ geographic background and comfort level with computer-mediated communication moderate the effectiveness of the course in terms of learning outcomes. Finally, qualitative data from the open-ended questions help support the quantitative analyses and provide a richer understanding of participants’ experiences and learning in the online course. Implications for research on the effectiveness of online diversity courses are discussed.