From a global perspective, climate change mitigation requires countries to consider Low-Carbon Development (LCD) policies, including “Low-Carbon Transport (LCT)” and decarbonization options, which should be ideally developed within a place-based scope. Due to the complex structure of the transportation sector, mitigation actions require not only technological but also behavioral shifts, which can be challenged by the existing legislative and socio-economic structure of a region. Therefore, it is essential to identify and assess barriers and opportunities (B&Os) as a necessary initial phase in the development of national mitigation strategies. Turkey voluntarily started climate change mitigation by preparing the legislative and political infrastructure as a part of the European Union Harmonization Process. Detection of B&Os in other dimensions required an in-depth evaluation of existing LCT barriers and instruments (B&Is) in the IPCC literature in terms of their applicability and priority for Turkey. A focus group study with transport sector representatives was conducted to evaluate 15 barriers and 19 instruments (proposing 285 B&I matrix combinations), followed by a brain-writing exercise that revealed 12 main barriers (i.e., a lack of measurable targets, an imbalanced modal split, a lack of institutional coordination and data archiving, etc.). Various opportunities have been identified, such as the increased utilization of Intelligent Transportation Systems and the timely adoption of vehicle and fuel technologies in the EU region. These opportunities would not only contribute to the development of a national climate change mitigation strategy but also facilitate the implementation of place-based action plans, including climate change action plans and sustainable urban mobility plans at the city level.