The purpose of this study was to investigate high school students' knowledge and attitudes regarding biotechnology and its various applications. In addition, whether students' knowledge and attitudes differed according to age and gender were also explored. The Biotechnology Knowledge Questionnaire (BKQ) with 16 items and the Biotechnology Attitude Questionnaire (BAQ) with 37 items which were adapted into Turkish were administrated to 352 high school students. The data were analyzed by making use of both inferential and descriptive statistics. The results showed that students attained poor knowledge regarding biotechnology and its various applications. We found no statistically significant difference between male and female high school students with regard to biotechnology knowledge. On the other hand, we found significant impact of age on biotechnology knowledge. The effects of gender and age were found significant on attitudes. Furthermore, a significant difference was observed between male and female students with regard to attitudes toward biotechnology and its applications in favor of males. The older the students were the more positive attitudes they had.