To foster sustainable regional development, many regions rely on innovations. To safeguard the generation of innovations and their market introduction, companies have increasingly used technology roadmapping and open innovation. The project INNOrural (Innovations for sustainable rural development) expanded these concepts by applying them to regions. This led to the rise of the “Regional Open Innovation Roadmapping” framework for innovation-based regional development (ROIR). This framework was tested by conducting two innovation roadmapping processes in the model region of Märkisch-Oderland (MOL), Germany: the certification of regional wood fuel and the establishment of a competence center for precision farming technology. Both innovation ideas were selected during the roadmapping process by applying a sustainability assessment. After 12 months, two complete roadmaps were ready for implementation. Key principles of ROIR were identified, including the use of a clear and replicable sustainability assessment method, the involvement of all relevant stakeholder groups in the early process and the cooperation between regional and subject experts. Generally, the broader adaptation of ROIR for additional regions will be useful. Nevertheless, the ROIR processes need to be evaluated in depth to develop a better understanding and to provide evidence of the benefits and limitations of this approach.