Historically, market regulation has played an important role in shaping the trajectory of scientific and technological innovation in food and agriculture. However, regulators’ traditional focus on safety and efficacy may be insufficient to address more complex ethical, legal, and social implications (ELSI) of novel products, such as the use of nanotechnology and nanomaterials in food and agriculture (nano-agrifoods). One solution might be to implement the principles of responsible innovation (RI) to challenge innovators and policymakers to better anticipate risks further upstream and be responsive to societal desires and concerns, although substantial barriers to implementation persist. This paper presents stakeholder views on the relationship between regulation and RI in nano-agrifoods based on a broader U.S. stakeholder engagement study conducted in the fall of 2020. We found that participants raised key issues that incorporated all 4 pillars of RI (anticipation, inclusion, reflexivity, responsiveness). We also found that participants’ attitudes about the relationship between regulation and innovation informed their recommendations about the relationship between regulation and RI. These attitudes are represented in a spectrum of views, ranging from “regulation as barrier” to “regulation as driver” of innovation. We further identified implications for how each attitude might be used to operationalize RI in regulatory systems. Overall, these results suggest that just as regulation drove key innovations in the twentieth century, regulation may still have a role to play in helping to promote RI of nano-agrifoods in the twenty-first.