While research suggests that the inclusion of students with Down syndrome (DS) in regular classrooms has a positive impact on their learning, in general, a supportive teacher with a positive attitude is fundamental in shaping these students' school experiences. In Ghana, despite a large number of children with DS, there is little research exploring preservice teachers' attitudes toward including students with DS in regular schools. In this study, we recruited preservice teachers from teacher training institutions to understand the predictors of their attitudes toward teaching students with DS in regular classrooms. Using the revised Sentiment, Attitude, and Concern about Inclusive Education scale (SACIE-R), 855 preservice teachers were recruited from three colleges of education (n = 524) and two campuses of a public university (n = 331). Although the preservice teachers were ambivalent about teaching students with DS, several demographic variables-such as preservice training in inclusive education, practice experience, and confidence teaching students with disabilities-emerged as significant predictors of attitudes. The study limitations, recommendations for future research and implications for policymaking are discussed.