Among ctenid spiders, ctenines comprise the most diverse subfamily. In this study, a new genus of Cteninae, Spinoctenus, is proposed to include the type species S. yotoco, sp. nov. Ten new species are also described: S. escalerete, S. pericos, S. eberhardi, S. spinosus, S. stephaniae, S. nambi, S. florezi, S. tequendama, S. chocoensis and S. flammigerus. Results of the parsimony and Bayesian phylogenetic analyses using morphological and behavioural characters indicate the monophyly of this genus, closely related to Phoneutria Perty, 1883 and Ctenus Walckenaer, 1805. This genus can be distinguished from the remaining Ctenidae by three unambiguous synapomorphies: embolus with folded process, tegulum with median process, and RTA curved internally close to the cymbium. A dispersal-vicariance biogeographical analysis of the genus in the Andean and Chocó regions indicates the origin of Spinoctenus in the Western and Central Andean Cordilleras. From this region, three events of dispersal occurred to the other regions (one to the Chocó and two to the Eastern Cordillera), which were subsequently followed by three events of vicariance, suggesting that dispersal and vicariance were equally important in shaping the current distribution patterns of Spinoctenus species. The discovery of this new genus containing a large number of new species in the Andean and Chocó regions highlights the current poor knowledge of the Colombian biodiversity.