Biotechnology presence both in curricula and among researches regarding its teaching and learning processes is constantly increasing. The knowledge, attitudes and interests of 184 Spanish pre-university students on biotechnology in the fields of health, food, industry and environment have been analyzed in this work. The results showed that while some areas are well known, favourable attitudes and interests depend on other factors. Thus, positive correlations have been found between: (1) knowledge and interest towards almost all areas considered (rs 0.152-0.214) except for fermentation, (2) knowledge and attitude regarding the use of fermentation for food production (rs 0.325) and applications related to improve the environment (rs 0.239), (3) and attitude and interest only in the medical field (rs 0.327). As a conclusion, it is important to consider the relationships between these three constructs for designing activities including socio-scientific issue in order to facilitate the analysis of information in a critical way.