Computer vision syndrome has been an issue of concern among students who use digital devices continuously. This study aimed to determine the prevalence and level of knowledge on computer vision syndrome and its relationship with associated factors among undergraduate students in a public university in Malaysia. This study was conducted between 26 May and 23 June 2022 at National University of Malaysia. A cross-sectional study among 208 undergraduate students from a public university was conducted. A self-reported questionnaire via Google Form was used to capture the data among the undergraduates. The prevalence and associated factors of computer vision syndrome were each evaluated using the validated Computer Vision Syndrome Questionnaire and Computer Vision Syndrome Survey Form 3 questionnaires, respectively, while knowledge of computer vision syndrome was assessed using a validated questionnaire from a previous study. All the data were analyzed using the Statistical Package for Social Sciences version 26.0 software (IBM Corp.; Armonk, NY, USA). The prevalence of computer vision syndrome among undergraduates was 63.0% (n=131), with 91.9% having poor knowledge of computer vision syndrome. Significant associations toward computer vision syndrome were found among undergraduates who have refractive errors/wearing glass (69.3%), screen edge at or above horizontal eye level (79.4%), uncomfortable sitting postures (79.4%) and close eye-screen distance (82.0%). In-depth analysis showed that having refractive errors/wearing glasses (aOR: 1.93; CI: 1.05, 3.57), uncomfortable sitting postures (aOR: 2.01; CI: 1.08, 3.74), and close eye-screen distance (aOR: 2.81; CI: 1.31, 6.05) had odd chance to develop computer vision syndrome. The study's findings denoted that digital device users should have more knowledge of computer vision syndrome and practice the preventable measures, such as proper viewing distance and angle, upright sitting postures, appropriate screen and surrounding illuminance, as well as regular eye check-ups.