We revised the taxonomic status of the putative Acomys spinosissimus complex based on the comparative study of speci-men collections from Tanzania, Zambia, Zimbabwe, Mozambique, DR Congo and South Africa, by means of analysis of external morphology, craniometry, enzymes, mitochondrial DNA sequences and karyology. Our results confirm that A. spinosissimus represents a complex of species with seemingly non-overlapping distribution ranges. The distribution range of A. spinosissimus appears to be restricted between the Zambesi and Limpopo Rivers, while the reinstated A. selousi (that includes A. transvaalensis) occurs further to the South (i.e. northern limit seemingly just north of the Limpopo River). The investigated populations north of the Zambezi River are morphologically and genetically distinct from A. spinosissimus and A. selousi. Based on this evidence, we described Acomys muzei sp. nov. and Acomys ngurui sp. nov., each one occur-ring separately along one side of the Eastern Arc Mountains. Finally, we lacked sufficient information to describe a third new species from the area north of the Zambesi River.