Food safety is a persistent global health challenge. Improper handling of foods increases the chances of food contamination, which may endanger the consumers’ health. The knowledge gap and improper hygienic practices of food handlers are the key factors for the transmission of foodborne diseases. This study aimed to investigate the level of food safety knowledge, attitudes, and practices among street food vendors in Dhaka City, Bangladesh. In this cross-sectional study, a randomly selected 350 street vendors from seven locations in Dhaka City were enrolled. Data were collected by trained interviewers using a questionnaire. Only 36.5% of the respondents had primary, and 48% had secondary education. The majority of the vendors who handled food in front of observers showed good handwashing practice. However, very few vendors showed positive attitudes for washing hands before handling raw or cooked food (3.7%), practicing good personal hygiene (1.2%), and other food safety attitudes (less than 1%). This study showed a significant correlation between the education level of the participants, work experience, and qualification of the head chef with knowledge about hand washing, wearing gloves, and food-borne illnesses. The study demonstrated that formal education played a significant role in vendors’ knowledge and practice of health safety measures of food handlers in preventing foodborne illness.