Effective biosecurity systems are essential to protect and add value to the world’s marine environments, and their associated commercial, cultural, and recreational values. The Marine Biosecurity Toolbox is a collaborative research programme whose mission is the development of science-based tools and technologies that empower governments, tangata whenua, industry and the public to effectively mitigate biosecurity risks. To realise this mission, the programme will develop a range of ‘tools’ that proactively prevent pest establishment, allow timely detection of incursions, and inform effective response strategies. Once implemented, these tools will enable a robust biosecurity system that adapts to future challenges and presents multiple barriers to the establishment and spread of marine non-indigenous species.