North Carolina’s extensive road network poses a serious risk to wildlife and disrupts the habitat connectivity that species need to move and survive. Collisions between larger wildlife and vehicles are also a constant human safety risk to travelers on North Carolina’s roadways. Wildlife road crossings are a proven method for solving both problems by allowing safe passage for wildlife and reducing wildlife-vehicle collisions. However, wildlife crossing structures are expensive, and even with substantial new federal funding sources coming online in recent years, there is still a pressing need to identify and rank the highest-priority locations in each state with the greatest need of these structures. Wildlands Network conducted a statewide project to identify hotspots for wildlife crossing need and highlight the top 20 highest-priority sites across North Carolina. We first identified over 150 draft sites using a combination of data sources including a geospatial prioritization model based on road segment and adjacent landscape characteristics, habitat connectivity networks at various scales, and an extensive set of wildlife-vehicle collision data from 2016-2020. Then, we surveyed wildlife experts across the state, providing them with a map of the draft sites and asking them to vote for the highest-priority locations (and suggest additional sites). The survey yielded 42 responses, and based on those votes we stratified our final top-20 sites across the four ecoregions of North Carolina to represent the state evenly. The results are presented in our new report “Prioritizing Wildlife Road Crossings in North Carolina,” which includes the top-20 sites and the full list of 179 hotspots. This project was designed to support competitive grant applications for federal wildlife funding, directing those efforts to the places in North Carolina with the most urgent need of wildlife crossing improvements. We also had the goal of supporting land conservation strategies to ensure long term wildlife connectivity at these sites, to help justify state DOT investments in infrastructure. Moving forward, we will work with NC DOT to ensure the report is integrated into the state’s ongoing highway improvement planning process (STIP) while also highlighting locations where stand-alone installations of wildlife crossings are justified by the significant risks to wildlife and motorists alike.