<p>The purpose of this paper is to conduct a systematic literature review on regional smart specialisation strategies, through a bibliometric and systematic analysis, combining performance analysis and scientific mapping to detect and visualise the concurrent subdomains and their evolution over time, pointing out future lines of research. The methodology of this systematic review is based on three phases: (1) database search, (2) bibliometric analysis, and (3) systematic analysis. In the first phase, the PRISMA protocol is used and publications are obtained from the Web of Science and Scopus databases. In the bibliometric analysis, the performance analysis and scientific mapping are carried out, using the R Studio software (version 4.02), using the biblioshiny with the bibliometrix package. The systematic analysis is based not only on the results obtained in the bibliometric analysis, but also on the methodological and content information taken from the publications. This article analyses and relates four clusters—knowledge, policy, innovation, specialization—indicated by the strategic diagram and trend topics, and 15 ‘themes’: policy, innovation, knowledge, priorities, systems, institutions, entrepreneurial discovery process, diversification, related variety, clusters, multi-level governance, helix model, competitive advantage, geography, and six critical questions. The paper systematizes the literature, filling a gap on the experience gained from the implementation of these strategies in the regional, addressing the critical questions and challenges that have arisen during their implementation in this context.</p>