USF may refer to:
- Universities
- University of South Florida, Tampa, Florida
- University of San Francisco, California
- University of St. Francis, Joliet, Illinois
- University of Sioux Falls, South Dakota
- Locations
- Universal Studios Florida, a theme park in Orlando, Florida
- File formats
- Universal Subtitle Format
- Nintendo Ultra 64 Sound Format
- Other uses
- Universal Service Fund, a United States government program
- Upstream stimulatory factor, a DNA-binding protein regulating gene expression