Greater Sage-Grouse (Centrocercus urophasianus) is a species of conservation concern and is highly susceptible to mortality from West Nile virus (WNV). Culex tarsalis, a mosquito species, is the suspected primary vector for transmitting WNV to sage-grouse. We captured, radio-tagged, and monitored female sage-grouse to estimate breeding season (April 15 to September 15) survival, 2016–2017. Deceased sage-grouse were tested for active WNV; live-captured and hunter-harvested sage-grouse were tested for WNV antibody titers. Additionally, we trapped mosquitoes with CO₂-baited traps 4 nights per week (542 trap nights) to estimate WNV minimum infection rate (MIR). Eight sage-grouse mortalities occurred during the WNV seasons of 2016 and 2017, 5 had recoverable tissue, and 1 of 5 tested positive for WNV infection. Survival varied temporally with sage-grouse biological seasons, not WNV seasonality. Survival was 0.68 (95% CI: 0.56–0.78; n = 74) during the reproductive season (April 1 to September 15). Mammalian predators were the leading suspected cause of mortality (40%), followed by unknown cause (25%), avian predation (15%), unknown predation (15%), and WNV (5%). These results indicate WNV was not a significant driver of adult sage-grouse survival during this study. Three sage-grouse (1.9%; 95% CI: 0.5–5.9%) contained WNV antibodies. We captured 12,472 mosquitoes of which 3,933 (32%) were C. tarsalis. The estimated WNV MIR of C. tarsalis during 2016 and 2017 was 3.3 and 1.6, respectively. Our results suggest sage-grouse in South Dakota have limited exposure to WNV, and WNV was not a significant source of sage-grouse mortality in South Dakota during 2016 and 2017. Based on our finding that a majority of sage-grouse in South Dakota are susceptible to WNV infection, WNV could potentially have an impact on the population during an epizootic event; however, when WNV is at or near-endemic levels, it appears to have little impact on sage-grouse survival.