As Nepal gradually integrates sexuality education into its formal school curriculum, understanding university students' knowledge and attitudes toward sex education is critical. Despite existing reforms, gaps remain in the delivery and reception of sexuality education at higher education levels. This study aimed to assess the level of knowledge and explore the attitudes of university-level students toward sexuality education in the Nepali context. A descriptive quantitative design was used to conduct a census-based study among 60 Bachelor’s and Master’s level Health Education (HPE) students at the Central Campus of Education, Mid-Western University. Structured questionnaires and interview schedules were used to collect data. Descriptive statistics (frequencies and percentages) were used to analyze student responses regarding knowledge and attitudes toward sexuality education. The study found that 96.67% of respondents were knowledgeable about sexuality education, with textbooks (61.67%) being the primary source of information. A majority understood the importance of sex education and were interested in learning more, particularly in behavioral and problem-solving aspects. Although 53.3% believed that teachers should teach students of the same gender, most rejected traditional taboos and misconceptions, such as equating sex education with pornography or considering sex a private matter. Attitudes toward harmful practices like Chhaugoth were also largely negative, indicating shifting norms. The findings suggest growing awareness and a positive shift in attitudes toward sex education among Nepali university students. However, cultural sensitivities, gender norms, and uneven knowledge of sexuality subtopics reveal the need for improved, inclusive, and culturally appropriate sex education programs. Structured teacher training and curriculum enhancements are recommended to address these gaps and support holistic adolescent development.