Understanding the human journey of global colonisation is the history of modern humanity and the development of the diverse characteristics of peoples and cultures around the world. This five-year interdisciplinary project will investigate the peopling of South America, the last continental terra incognita (other than Antarctica) to be colonised by humans, constituting a virtually unprecedented migration of modern humans across richly diverse, empty landscapes during the Late Pleistocene-Early Holocene transition. Situated at the geographical gateway to the continent, the project will investigate one of the most momentous demographic dispersals of our species into the diverse environments of north-western South America, encompassing coasts, savannahs and lowland, Sub Andean and Andean tropical forests. This process took place amidst one of the most significant climatic, environmental, and subsistence regime shifts in human history, which contributed to the extinction of megafauna, plant domestication, and today’s remarkable diversity of indigenous South American groups. Despite its geographical importance and a wealth of archaeological and palaeoecological data across its diverse environments, north-western South America has only been given cursory consideration to understand processes of human dispersion. This project will redress this imbalance by applying an innovative interdisciplinary approach that integrates state-of-art archaeology, archaeobotany, zooarchaeology, palaeoclimatology, palaeoecology, ancient environmental DNA and isotope studies. The results will provide a global comparative perspective to the study of Late Pleistocene human colonisations, hunter-gatherer adaptations, the demise of megafauna and the beginning of plant cultivation and domestication. The results of the project have broader implications not only for archaeology but also for geography, palaeoclimate, palaeoecology, and molecular biology.