As part of the Convention on Biological Diversity (CBD), the Nagoya Protocol on Access and Benefit Sharing calls for ‘fair and equitable sharing of benefits’ derived from the use of genetic resources and traditional knowledge. However, implementation of the Convention and the Nagoya Protocol has been challenged by the inadequacies of existing policies, lack of national-level frameworks, and inadequate knowledge among stakeholders. We used focus group meetings and structured interviews with rural communities, government representatives, researchers and Members of Parliament in Bhutan to collect data on awareness, knowledge and perceptions of components of the CBD related to access and benefit sharing. Our study indicated generally low levels of awareness about most components of the Convention, particularly among rural residents. Although local people in rural communities feel that benefits derived from local biological resources and traditional knowledge should be shared, there is uncertainty about who owns these resources. These results indicate that there is an urgent need to develop educational and awareness programmes, using a variety of media, to target particular stakeholder groups, with emphasis on residents in rural communities. This could empower local communities to participate meaningfully in decision-making processes to develop Bhutan's national access and benefit sharing framework, and to allow them to benefit from the conservation and sustainable use of local resources.