Mammogram screening (MS) is the gold-standard method for early detection of breast cancer (BC), and its use has been proven to minimize BC-related deaths and reduce treatment costs. However, recent epidemiological surveys have reported that rates of mammogram uptake by the Saudi female population are low. Here, we assessed the knowledge of BC and perceived barriers to MS uptake among pre-eligible northern Saudi women. We administered a standard and validated Arabic questionnaire to 400 women aged 40-69 years. SPSS version 21.0 (IBM Corporation, Armonk, NY, USA) was used for data analysis. We applied logistic regression analysis to find the factors associated with participants' knowledge of BC and MS. Spearman correlation test was applied to find the correlation between knowledge and barrier scores. The study participants reported that smoking habits (61.3%) and unhealthy food habits (57.8%) were the most common risk factors for BC. Of the studied participants, 56.3% had low or medium degrees of knowledge about BC risk factors and MS. The degree of knowledge was significantly associated with education level (adjusted odds ratio [aOR], 2.35; 95% confidence interval [CI]. = 1.61-3.13; P = 0.008) and a family history of BC (aOR, 3.66; 95% CI, 1.94-5.49; P < 0.001). Fear of a BC diagnosis (50.8%) and concerns regarding test procedures were the most common barriers to MS uptake. We also found a negative correlation between participants' knowledge and perceived barriers to MS (rho = -0.389, P < 0.001). We recommend that concerned authorities offer women multiple health education sessions covering BC risk factors and the necessity for pre-eligible women to undergo MS spaced at regular intervals at different facilities. Furthermore, a multicentric mixed-methods survey is warranted to find the qualitative aspects of barriers to MS.