Knowledge transfer involves the incorporation of knowledge into the practices of social and collective learning processes. This transfer occurs through social practices (Von Krogh, The Journal of Strategic Information Systems 21:154–164, 2012) and both tacit and explicit means (Nonaka and Takeuchi, The knowledge creating company: How Japanese companies create the dynamics of innovation. Oxford University Press, 1995), involving both individual and organizational actors. Universities contribute to this process through the “third mission” (Centobelli et al., Management Decision 57:3301–3320, 2019; Croce et al., S, The Journal of Technology Transfer 39:688–715, 2014; Paoloni et al., Business Process Management Journal 25:185–201, 2019), which includes scientific, technological, and cultural transfer and productive transformations of knowledge. This transformation finds its expression in the establishment of start-ups. However, despite efforts, a gender gap persists in the presence of women-led start-ups in Italy (Modaffari et al., Journal of Intellectual Capital 24:1506–1531, 2023; Modaffari and della Corte, Organizational resilience and female entrepreneurship during crises: Emerging evidence and future agenda. Springer, 2022; Modaffari and Manzo, ICGR 2022 5th international conference on gender research, Academic Conferences and Publishing Limited, 2022; When the crisis becomes an opportunity: The role of women in the post-covid organization (pp. 75–87), Springer, 2023). This study aims to investigate whether state universities in the Lazio region offer specific incentives for women-led start-ups. To achieve this goal, a multi-case qualitative analysis was conducted. The case was developed by first analyzing the guidelines issued by the universities to verify if they offer incentives for the establishment of women-led start-ups, and subsequently, the composition of active start-ups was analyzed to investigate whether these incentives contribute to increasing the number of women-led start-ups. The results highlight that, despite universities increasingly contributing to knowledge transfer, the gender gap remains strong. Despite the limitations imposed by the choice of universities to analyze, this study has the potential to enhance existing literature in gender studies and entrepreneurial universities, with particular emphasis on the domain of the third mission.