Objective The aim of this study was to assess COVID-19-related knowledge, attitudes, and preventive practices of two Pakistani university populations. Methods A cross-sectional study was conducted among students and employees of two higher education institutions in Lahore, Pakistan, namely the University of Lahore and the Gulab Devi Educational Complex. Participants were recruited using a convenient sampling method. A validated 45-item, self-administered questionnaire was used to collect data. Total possible scores were 0-14 for knowledge (scores < 7, 7-10 and > 10 were considered to indicate poor, moderate and good knowledge, respectively); 0-7 for attitude (scores > 5 were considered to indicate a good attitude); and 0-18 for COVID-19 preventative practices (scores > 12 were considered to indicate good preventative practices). Results Of the 417 enrolled participants, 416 reported that they were aware of COVID-19 and social media was the major source of their information. Mean scores were 10.12 ± 2.20 for knowledge (good, moderate and poor knowledge in 50.2%, 42.8% and 7.0% of participants, respectively); 5.74 ± 1.28 for attitude (65.4% of individuals had a positive attitude); and 11.04 ± 3.34 for COVID-19 preventative practices (only 36.5% of participants had good preventive practices). Conclusion Pakistani university students and employees have good knowledge and attitudes regarding COVID-19, but unsatisfactory preventive practices. Therefore, health regulators should use multiple communication approaches, such as electronic, print and social media, phone messages, etc., to increase awareness and improve practices related to COVID-19. I use a face mask in the crowds and when I visit healthcare settings nowadays 148 (35.6) 88 (21.2) 104 (25.0) 76 (18.3)