Abstract: As human development continues to expand, wildlife must relocate or adapt to survive. Many mammalian mesopredators, such as the Virginia opossum (Didelphis virginiana), have adapted to living alongside human development. Furthermore, top‐down predation pressure may be altered in nuanced ways within the human environment. Species such as opossums may be shielded from predation by human development or behavioral changes in predators. Understanding how dominant and subordinate mesopredators co‐exist across natural and developed areas will provide insight into how wildlife communities are structured. Our objective was to evaluate how opossum occupancy, abundance, and activity were associated with human development and the relative abundance of their predators. We used data from a nationwide camera trapping study, Snapshot USA, to estimate opossum occupancy, abundance, and activity. We related these measures to the surrounding landscape and urbanization variables. We found that opossum occupancy was positively associated with anthropogenic sound (a surrogate for human activity). Furthermore, opossums in heavily forested areas were more likely to be detected in locations with higher predicted anthropogenic sounds. In areas with a high density of human housing, opossum relative abundance increased when predator abundance increased. We also found opossums were strictly nocturnal and shifted their activity to earlier in the evening in the presence of high predator abundance. Our results suggest that humans and their urban development can have multidimensional impacts on opossum behavior and occurrence, and could facilitate changes in predator–prey dynamics. Future research should evaluate if the association of opossums with urban areas is due to human‐subsidized resources or caused by reduced mortality from altered predator–prey dynamics.