Information security awareness (ISA) has become a vital issue, as security breaches often attributed to humans lead to losses for individuals and organizations. Information security (IS) education may be an effective strategy to improve students’ ISA; however, studies associated with the relationships between teaching effects and information security learning are few. This study adopted gamification practice and examined its effect on students’ ISA knowledge enhancement, attitude and intention of security compliance, and willingness for continuous IS education. This study also examined the gender difference in a gamified learning system. One hundred ten undergraduates participated in a quasi-experimental study. The results indicated that students within a gamified class performed better than students within a lecture-based instructional group. We found significant gamification effects on the three security focus areas of password management, Internet use, and information handling. Gamification did not significantly impact the attitude and intention of participants’ security compliance and students’ willingness for continuous IS learning. Gender difference in the effect of gamification on ISA knowledge enhancement was not observed as well. The research provides theoretical and practical contributions by incorporating gamification into IS learning and suggests gamification as an effective means to enhance students’ knowledge acquisition in an engaging, timely, economical, and repeated manner.