This paper investigates the level of knowledge and interest in biotechnology education of Malaysian secondary school students. The research was based on a questionnaire adapted from the instruments developed by Prokop et al. and Kidman. Six schools in the Klang Valley were involved in the research and with participation by a total of 427 Grade 11 students (aged 17 years). The findings showed that the students possess a moderate level of biotechnology knowledge in all the five components assessed. The five components include Genes and Chromosomes, DNA Fingerprinting, Genetic Engineering, Human Genome Project, and Stem Cell Research. In addition, results revealed that students are interested to find out more about the adverse effects of genetic engineering in the field of medicine, consumption of genetically modified foods, and on the environment.