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Background: Even though organ transplants save thousands of lives globally, there exists a wide mismatch between an increasing demand for organ transplants and limited number of organ donors. The aim of the study was to assess the reasons and inspect differences between donors and non-donors in both students of medical and non-medical universities. Methods: A cross-sectional study was conducted over a period of 6 months (May 2018-March 2021). Sample of 400 participants enrolled from three medical colleges, two business/social sciences institutions and one engineering university of Karachi. Categorical variables were compared using the Chi-squared test. Continuous variables were compared using a student t-test. Variables with a univariate p-value ≤ 0.10 were included in multiple logistic regression models. Adjusted odds ratio (OR) with 95% confidence interval were reported. Results: With a very high response rate 383(96%) for knowledge about organ donation, a subdued response 201(52.3%) for willingness to donate. Most common source of knowledge regarding organ donation was social media 249(65%), followed by university 207(54%), family/friends 184(48%) and television 172(45%). Multiple logistic regression model identified being comfortable discussing organ donation as a positive predictor of willingness to donate (OR: 2.71, p=0.005), and the belief that organ donation disfigures the body as a negative predictor (OR: 0.40, p