Genetic diversity is theorized to decrease in populations closer to a species' range edge, where habitat may be suboptimal. Generalist species capable of long-range dispersal may maintain sufficient gene flow to counteract this, though the presence of significant barriers to dispersal (e.g., large water bodies, human-dominated landscapes) may still lead to, and exacerbate, the edge effect. We used microsatellite data for 2421 gray wolves (Canis lupus) from 24 subpopulations (groups) to model how allelic richness and expected heterozygosity varied with mainland–island position and two measures of range edge (latitude and distance from range center) across >7.3 million km² of northern North America. We expected low genetic diversity both at high latitudes, due to harsh environmental conditions, and on islands, but no change in diversity with distance to the range center due to the species' exceptional dispersal ability and favorable conditions in far eastern and western habitats. We found that allelic richness and expected heterozygosity of island groups were measurably less than that of mainland groups, and that these differences increased with the island's distance to the species' range center in the study area. Our results demonstrate how multiple axes of geographic isolation (distance from range center and island habitation) can act synergistically to erode the genetic diversity of wide-ranging terrestrial vertebrate populations despite the counteracting influence of long-range dispersal ability. These findings emphasize how geographic isolation is a potential threat to the genetic diversity and viability of terrestrial vertebrate populations even among species capable of long-range dispersal.