With only five protected areas dedicated to the conservation of biodiversity (two national parks, one strict nature reserve and two faunal reserves), Guinea has one of the smallest protected area networks in West Africa. As a result, two of the five ecoregions of the country and six of the 14 globally threatened large and medium-sized mammals occurring in Guinea are not found in the national protected area network. To identify areas with high biodiversity that could be included in the national protected area network, we used the Key Biodiversity Areas (KBA) methodology. We devised a scoring system to rank the identified KBAs according to their relative conservation significance. We identified a total of 16 KBAs throughout the country. Their proclamation as protected areas would result in the protection of all ecoregions and all but one of Guinea's globally threatened large and medium-sized mammals. Twelve of the 16 KBAs have the legal status of classified forest, a status that should facilitate the change into formal biodiversity protected areas (IUCN category I-IV). Our analysis indicates that even if only the two areas with the highest conservation significance score, the Ziama and Diécké forests, become formal protected areas, this would provide protection to both the western Guinean lowland forests, one of the most threatened ecoregions in Africa, and to 11 of the 14 threatened large and medium-sized mammals occurring in Guinea.