This cross-sectional study comprises a questionnaire-based survey regarding knowledge about human papillomavirus and its vaccine among students in different educational fields at public and private universities in the city of Lahore in Pakistan. A 26-item questionnaire was used to attain the objective of this study. The reliability of this tool was assessed using Cronbach’s alpha (0.79) and the Kaiser-Meyer-Olkin value was 0.827. The response rate to the survey was 78.0%, of whom the majority (74.9%) were females and 308 (79%) were single (median age = 23 years). While assessing the respondents’ knowledge about HPV, 223(57%) students reported that they had already heard of HPV (human papillomavirus) and nearly 215 (55%) reported that HPV causes cervical cancer and can infect both men and women. Gender and field of study were two main factors found influencing the respondents’ knowledge about HPV. Moreover, students’ understanding about the mode of transmission of HPV was cursory: 40.51% said they didn’t know how HPV is transmitted, 133 (34.10%) stated that HPV spreads through the exchange of bodily fluids, and 22 (5.64%) selected cough/sneezing. In terms of prevention, 175 (44.87%) students stated that HPV can be prevented by vaccination, 30.0% reported sexual abstinence, 21.54% using condoms, and nearly 5.38% disclosed use of antibiotics. Addressing the knowledge of students regarding HPV vaccine, nearly 53% stated there is no vaccine against HPV and almost 64% rejected the statement that HPV vaccine prevents cervical cancer. In addition, students reported that they will be more than willing to get vaccinated for HPV if their physician recommend them (RII=0.74) followed by parents (RII=0.69). The results of this study revealed a poor understanding among respondents about the health problems associated with HPV, its prevention, modes of transmission and arability of HPV vaccine in Pakistan.