Research related to the teaching and learning of 21st century biotechnology in additional science in Malaysia is seldom conducted. This study was carried out to find out the problems teachers and students face in biotechnology teaching and learning, and the appropriateness of the inclusion of 21st century skills, namely digital-era literacy skill, inventive skill, and spiritual values. A survey was conducted in Malaysian secondary schools aimed at students aged between 16 and 17, and teachers. Results showed that the inclusion of 21st century skills in biotechnology was relevant, and recommendations were given by the participants to improve the teaching and learning of biotechnology that could further cultivate their scientific literacy, especially in biotechnology.