One of the most important opportunities in the United States for mitigating highway effects on wildlife is in the Pigeon River Gorge area where Interstate 40 winds through the mountains of NC and TN, cutting off the tremendous biodiversity of Great Smoky Mountains National Park from extensive national forest lands to the northeast. This region is home to abundant large-bodied wildlife species such as elk, black bear, and white-tailed deer, in addition to an incredible range of smaller species ranging from snakes to bobcats to salamanders. Given the ecological diversity and importance of the region and the severity of elk- and bear- vehicle collisions, conservationists are seeking ways to alleviate roadway barrier effects, restore connectivity, and protect wildlife and human safety. With support from state, federal, and NGO partners, our multifaceted field research approach focuses on a 28-mile section of I-40 where we've been conducting research since 2018 to meet the following objectives: (1) evaluate existing levels of roadway permeability by documenting wildlife use of 20 structures (i.e., culverts, bridges, underpasses) with camera traps (n=40), (2) detect areas of relatively high roadside animal activity with camera traps (n=68) placed within the forested right-of-way of a stratified sample of road segments, (3) identify locations with high incidences of wildlife road mortality through weekly driving surveys and data-sharing agreements with state DOTs and other agency partners, and (4) identify and predict elk road conflict locations using fixes from 13 GPS-collared elk and subsequent geospatial analyses. We will present findings from our research and demonstrate how this research will provide the framework that identifies areas where mitigation strategies such as road crossing structures could be best implemented along the interstate to reduce wildlife vehicle collisions and increase wildlife habitat connectivity.