There are 17 players on the 2001 Project-40 roster:
| Player | Birthdate | Hometown | MLS Club |
| Shaker Asad | 8-18-79 | Raleigh, N.C. | New England |
| Devin Barclay | 4-9-83 | Anapolis, Md. | Draft Eligible |
| Isaias Bardales Jr. | 8-18-79 | San Jose, Cailf. | Draft Eligible |
| DaMarcus Beasley | 5-24-82 | Ft. Wayne, Ind | Chicago |
| Kyle Beckerman | 4-23-82 | Crofton, Md. | Miami |
| Scott Bower | 6-27-78 | Tampa, Fla | San Jose |
| Chris Carrieri | 4-28-80 | Stafford, Va. | Draft Eligible |
| Bobby Convey | 5-27-83 | Philadelphia, Pa. | D.C. United |
| Micah Cooks | 7-2-81 | Bethesda, MD | D.C. United |
| Francisco Gomez | 1-25-79 | Watsonville, Calif. | Kansas City |
| Michael Green | 7-16-78 | Columbia, Md. | Kansas City |
| Mario Longo | 12-3-80 | Raleigh, N.C. | Columbus |
| Marvin Quijano | 10-9-79 | N. Hollywood, Calif. | Los Angeles |
| Nick Rimando | 6-17-79 | Montclair, Calif. | Miami |
| Miguel Saavedra | 7-3-83 | Milwaukee, Wis | Draft Eligible |
| Sergio Salas | 3-21-81 | Falls Church, Va. | D.C. United |
| Seth Trembly | 3-21-82 | Littleton, Colo. | Colorado |