This research examines the evolution of the study on triple helix (TH) approaches to the investigation of regional innovation systems (RISs). Prior studies on TH-based RISs have focused largely on complex interactions among innovation creating actors (i.e., university, industry, and government) and non-market institutions in various regional contexts. Although there are a growing number of TH-based RIS studies, they often ignore the evolutionary aspect of the RIS studies and implications for sustainability. Thus, herein we conduct a wide range of systematic literature review on the TH research in the RIS literature published from 1990 to 2017. The study identifies that the TH literature evolves into various N-helix models in particular, gravitating toward stakeholders, internationalization, specialization and ecological conservation, which offers various implications for sustainability in the regional development context. In the ages of globalization, digitalization, and socio-ecology, this study offers a comprehensive view on the TH research for sustainable development and suggests useful future research directions that academics and policymakers can benefit from.