Background: There is a close link between an individual's knowledge about a given disease and uptake of screening and ultimately treatment. This study aimed to determine the link between knowledge levels and awareness and uptake of cervical cancer screening among women of reproductive age (15 to 49 years) in Dodoma, Tanzania. Methods: A cross-sectional study of 1,587 women aged between 15 and 49 years was conducted in Dodoma City, Tanzania. A structured questionnaire, adapted from Montgomery and others, was pretested and used to collect data from March to April, 2016 via multistage sampling. Univariate and multiple regression analyses were used to determine factors associated with the level of knowledge about cervical cancer and the association between knowledge and uptake of cervical cancer screening. Results: The mean age of the participants was 26.99±8.026 years. Only 165 (10.4%) of the 1,587 participants were knowledgeable about cervical cancer; 1,051 (66.2%) were aware of cervical cancer screening, and only 125 (7.9%) had undergone cervical cancer screening. Predictors of knowledge about cervical cancer were education level (secondary education adjusted odds ratio [AOR] 2.23; 95% confidence interval [CI], 1.030-4.811; P<.05; university level AOR 2.59; 95% CI, 1.179 to 5.669; P<.05); residence (rural AOR 1.85; 95% CI, 1.282 to 2.679; P=.001); parity (multipara AOR 1.88; 95% CI, 1.125 to 3.142; P<.05). After adjusting for confounders, knowledge about cervical cancer significantly influenced both cervical cancer screening awareness (AOR 2.91; 95% CI, 1.821 to 4.640; P<.001) and uptake (AOR 2.065; 95% CI, 1.238 to 3.444; P=.005). Conclusion: The level of knowledge about cervical cancer was extremely low. Women with less knowledge about cervical cancer were those with less education, those living in rural areas, and those without children. A low level of knowledge was associated with poor uptake of screening services, highlighting the need for integrating health education pertaining to cervical cancer and screening when providing reproductive health care in Tanzania.