Abstract The current study investigated the impact of roads on native wildlife fauna in Pothwar Plateau, Pakistan. By conducting driven transects over a period of 24 months (March 2012–February 2014), we identified 131 carcasses resulting from wildlife-vehicle collisions or “road kill”, comprising 18 vertebrate species from seven different orders. The Asiatic jackal (Canis aureus; Linnaeus, 1758) was the most common road kill (37.4%), followed by the small Indian mongoose (Urva auropunctatus; Hodgson, 1836; 9.9%) and the red fox (Vulpes vulpes; Linnaeus, 1758; 8.4%). Road kill incidences included both nocturnal (55.6%) and diurnal species (44.5%). The maximum number of road kills were encountered in the Rawalpindi district area (46.6%), followed by Chakwal (36.6%), Jhelum (8.4%) and Attock (8.4%). The majority of road kills occurred on paved roads (93.9%; 0.009 road kill/km) with significantly fewer occurrences on unpaved roads (6.1%; 0.004 road kill/km). Similarly, 77% (0.009/km) of the road kills were found adjacent to protected areas. The majority of road kills (33%) occurred during the spring (February–April) but less in summer (May–July) and winter (November–January) (19.8% for both). The study provides important data on road mortality of wildlife species in the region bordering protected areas in the Pothwar Plateau, Pakistan, to initiate management and conservation strategies.