Biotechnology holds immense promise for advancing sustainability across sectors such as healthcare, materials, chemicals, and food production. Yet, its adoption remains hindered by public scepticism, industry hesitation, and regulatory inertia. The B-trust project addresses this challenge by developing a transparent and inclusive governance model that fosters trust in biotechnology, particularly within the bio-based and agri-food sectors. Grounded in the Theory of Change, the project engages civil society and stakeholders through co-creation processes that serve as trust-building catalysts. These efforts culminate in a set of principles and measures designed to enhance transparency, safety, and stakeholder engagement. Six biotech use cases, selected to reflect diverse concerns and benefits, serve as the foundation for mapping real and perceived risks. This comprehensive approach enables a nuanced understanding of biotechnology’s societal impact and lays the groundwork for broader acceptance. A dedicated forum ensures ongoing training and stakeholder involvement, supporting the model’s replication and long-term sustainability in research, policy, and funding contexts.