To address the ongoing global biodiversity crisis, conservation approaches must be underpinned by robust information. Canada is uniquely positioned to contribute to meeting global biodiversity targets, with some of the world’s largest remaining intact ecosystems, and a commitment to co-application of Indigenous ways of knowing alongside scientific, socioeconomic, and other approaches. We elicited input from experts across a range of disciplines to identify the key information needed to advance policy and management actions to conserve biodiversity in Canada. Experts concluded that, in many cases, a lack of information is not the major barrier to biodiversity conservation; instead, mechanisms to translate information into action are most urgently needed. Recognizing multiple ways of knowing, especially Indigenous knowledge systems, will be critical to support the transformative change needed to conserve biodiversity at a national scale. Collaboration among natural, social, and data scientists can facilitate social change and biodiversity information management. Experts identified 50 priority information needs which emphasize the importance of (i) reviewing policies and actions and disseminating lessons learned from successes and failures; (ii) better understanding mechanisms to build public support; (iii) improving, in specific instances, understanding of the status and trends of habitats, species, ecosystems, and threats for planning and management; and (iv) mobilizing biodiversity information. Through the Convention on Biological Diversity, the global community has resolved to “live in harmony with nature”; through our Canadian case study, we conclude that the most pressing need to address this resolution is an improved understanding of how to move from conservation knowledge to conservation action.