The present study explored the relationship between undergraduate students’ digital literacy and self-regulation in online interaction (student-content, student-teacher, student-student). Investigating this relationship can facilitate identifying areas for improvement of support programmes and provide insights for effective online teaching and learning practices. The researchers collected data from 195 undergraduate students during the Covid-19 pandemic. Canonical correlation results indicated a significant and positive correlation (Rc = .41), meaning that having a positive attitude and high technical knowledge and skills regarding digital technologies can help students manage online teacher, peer, and content interactions. Moreover, the study identified that the digital literacy scale’s technical variable contributes to the correlation more than the attitude variable. This result implies that developing technical knowledge and skills might be more critical to promoting self-regulation in online interaction. Based on these results, the implications for online education and recommendations for supporting self-regulation in student interactions are discussed.