The Flint Hills represent the largest tract of tallgrass prairie in North America and is located near the western edge of the native range of the Virginia opossum (Didelphis virginiana). This region is undergoing rapid landscape changes (e.g., urbanization, agriculture, woody encroachment) that are negatively affecting mammal communities. Although previous research has revealed northward distributional expansions of Virginia opossums facilitated by urban development, no studies have assessed how landscape change affects distribution patterns along the western edge of their geographic range. During 2016–2018, we monitored site (n = 74) occupancy along urban–rural transects in the Flint Hills to assess the influence of landscape change (i.e., urban, grassland, agriculture, woody encroachment) and water availability on the distribution of Virginia opossums. Sites surrounded by urban land cover had greater initial occupancy probabilities and lower extinction rates. Sites closer to permanent water sources experienced greater colonization rates and lower extinction rates. In addition, site extinction rates were lower in areas surrounded by woody encroachment. Our results concur with other studies suggesting that growing urban areas may expand opossum distributions along the edges of their geographic range. Our study also suggests that woody encroachment into tallgrass prairies may provide an alternative pathway for future distributional expansions. Future research must consider the potential for landscape change, along with dynamic water availability, in models predicting the distribution of Virginia opossums.