The paper aims to examine the relationship between Entrepreneurial Orientation (EO), Market Orientation (MO) and performance in University Spin-Offs (USOs). Initially, the paper, assesses the existence of a moderating effect of MO in the relationship between EO and performance, next, it analyses the mediation role of MO between EO and performance. To strengthen paper results, we test our hypotheses on a cross-sectional sample of 162 Italian USOs using both perceived and objective performance. Findings suggest that there is no synergistic effect of MO and EO as mutually independent constructs. Instead, our evidences support the idea that EO and MO in USOs occur within the same learning process. Both EO and MO support USO performance, but MO cannot occur without EO as an antecedent condition. At the same time, a significant portion of the EO contribution to performance occurs through MO.