Background: Several studies reported that breast cancer is the most common cancer in women, and is the principal cause of cancer deaths and is therefore, a world concern. Early detection of breast cancer using breast self-examination (BSE) plays an important role in decreasing its morbidity and mortality. Objective:To assess the level of knowledge of breast cancer and practice of breast self-examination among female students in Rift Valley University. Methods and Materials:Cross-sectional study design was employed. A total of 423 female students were included. Stratified sampling method was used. Students were divided into two strata, as health science students and non-health science students. A pretested structured self-administered questionnaire was used to collect the data. Data was analyzed using SPSS version 20. Binary and Multiple logistic regressions were performed. Adjusted odds ratio (AOR) with 95% CI was used to measure the strength of association. Statistical significance was set at p<0.05Results: Most of the respondents (94.1%) were with age of 20-30 years. The mean score knowledge about breast cancer was 6.5. Out of 423 female students, 59.8% of them had a good knowledge about breast cancer. Half of them (54.4%) reported that they had heard about breast self-examination while 25.8% reported that they had practiced breast self-examination before. Students who had family history of breast cancer were more likely knowledgeable than who hadn't family history of breast cancer (COR=3.042, AOR=1.225, 95% CI=1.636-5.656).Conclusions: Nearly half of them had poor knowledge about breast cancer. Majority of them had poor knowledge how to perform breast self-examination and most of them didn't perform breast self-examination before.