Universities were identified as key-actors in the process of designing and implementing of S3 strategies (Goddard, J., Kempton, L., 2011). This paper will explore how the regional innovation system approach as an instrumental policy for growth is influencing universities transformation towards assuming the third mission. The main research question is if the active involvement of universities in S3 implementing creates effects on their entrepreneurial potential. I have applied a qualitative research, using a bibliometric research regarding universities role in regional innovation system. Also, in the framework of indicators proposed by Regional Innovation Scoreboard, I assessed the contribution of the universities from North-East region of Romania to regional innovation. The findings of this research are identifiying the main challenges universities are confronted with in order to contributing to regional development as part of their third mission.