The Proterozoic Athabasca Basin is well known for its unusually large-tonnage and high-grade 'unconformity-related' uranium (U) deposits, however, explanations for the basin-wide U endowment have not been clearly identified. Previous studies indicate that U-rich brines with up to ~600 ppm U and variable Na/Ca ratios (from Na-dominated to Ca-dominated) were present at the sites of U mineralization, but it is unknown whether such fluids were developed solely in the vicinity of the U deposits or at a basinal scale. Our microthermometric and LA-ICP-MS analyses of fluid inclusions in quartz overgrowths from the barren part of the basin indicate that U-rich brines (0.6 to 26.8 ppm U), including Na-dominated and Ca-dominated varieties, were widely developed in the basin. These U concentrations, although not as high as the highest found in the U deposits, are more than two orders of magnitude higher than most naturally occurring geologic fluids. The basin-scale development of U-rich diagenetic fluids is interpreted to be related to several geologic factors, including availability of basinal brines and U-rich lithologies, and a hydrogeologic framework that facilitated fluid circulation and U leaching. The combination of these favorable conditions is responsible for the U fertility of the Athabasca Basin.