Despite its extreme environmental conditions the deep sea harbors a unique and species-rich fauna of mostly unknown age and phylogeny. Asellote isopods have undergone their most impressive radiation in the deep sea, being found at all depths down to the deepest trench. Here we present the first molecular evidence for the phylogenetic origin of this remarkable array of deep-sea crustaceans, based on 30 new DNA-sequences of the complete 18s rRNA gene of specimens collected at depths down to 4543 m in the South Atlantic and South Polar Ocean. The results show that most of these isopod lineages belong to a single ancient clade. They evolved in situ in large oceanic depths and survived several climatic changes, but the lack of fossils and of a suitable molecular clock model prevents a precise dating of this radiation. The monophyly of typical deep-sea families, for example the Haploniscidae, Ischnomesidae or Munnopsidae, is well supported by different methods of analysis, while the monophyly of the Janiridae is rejected.