Teachers’ attitudes towards sexuality education are among the important predictors of their willingness to teach sexuality education programmes in schools. While there is a plethora of studies on teachers’ attitudes towards sexuality in developed countries, there is a paucity of such studies in sub-Saharan Africa in general and Tanzania in particular. This study examined teachers’ attitudes towards and comfort in teaching sexuality education in rural and urban Tanzania. The results show that an overwhelming majority of teachers in both rural and urban districts supported the teaching of sexuality education in schools, and the inclusion of a wide range of sexuality education topics in the curriculum. Nevertheless, though teachers expressed commitment to teaching sexuality education in schools, they expressed difficult and discomfort in teaching most of the key sexuality education topics. This implies that declaration of positive attitudes towards teaching sexuality education alone is not enough; there is a need for facilitating teachers with knowledge, skills and confidence to teach various sexuality education topics.