Background: Cervical cancer, one of the gynecological cancers, is a serious health concern in Ghana. To improve the chances of survival and treatment outcomes for this condition, early screening and detection is the best remedy. Comprehensive knowledge and positive attitude highly influence acceptability and uptake of screening methods. Objective: To assess the knowledge about cervical cancer and screening practice among females in secondary and tertiary institutions and uneducated females in Tamale-Ghana. Methods: Females (n = 300) between the ages of 15 and 49 years, comprising 100 participants in three categories (senior high school, tertiary institution and the non-educated) were recruited into the study. Data was gathered through a semi-structured questionnaire, and analyzed by descriptive statistical methods. Results: Approximately 61.3% of the participants have heard about cervical cancer, 33.1% and 29.9% of them got the information from school and the media respectively. Of those who have heard about cervical cancer, only 19.3% had considerable general knowledge about the condition. A significant association (p = 0.02) was observed between educational level and knowledge about cervical cancer. Meanwhile, only 5.3% of the 300 participants were previously screened for cervical cancer. Lack of information about cervical cancer was the most reported reason (46.6%) for not attending cervical cancer screening. Conclusion: Proactive intervention is required in the study area towards preventing new diagnosis through mass education, establishment of cervical cancer control programs, as well as screening and treatment centers.