The 2000–2001 Obie Awards, honoring distinguished achievement in Off-Broadway and Off-Off-Broadway productions, were presented on May 21, 2001, at New York's Webster Hall.
| Best Play: | Pamela Gien, The Syringa Tree |
| Playwriting: | José Rivera, References to Salvador Dalí Make Me Hot |
| Direction: | Michael Greif, Dogeaters; |
| Craig Lucas, Saved or Destroyed |
| Performance: | George Bartenieff, I Will Bear Witness; |
| Stephanie Berry, The Shaneequa Chronicles; |
| Ronnell Bey, Mandy Gonzalez, Judy Kuhn, and Anika Noni Rose, Eli's Comin'; |
| Bette Bourne, Resident Alien; |
| Brian d'Arcy James, The Good Thief; |
| Janie Dee, Comic Potential; |
| Jackie Hoffman, The Book of Liz; |
| Pamela Isaacs, Newyorkers; |
| Brian Murray, The Play About the Baby; |
| John Ortiz, References to Salvador Dalí Make Me Hot; |
| Mary-Louise Parker, Proof |
| Set Design: | Neil Patel, I Will Bear Witness, Race, Resident Alien, and War of the Worlds; |
| Douglas Stein, Saved and Texts for Nothing |
| Music: | Diedre Murray, vocal and instrumental arrangements, Eli's Comin'; |
| Bill Sims, Jr., Lackawanna Blues |
| Choreography: | John Carrafa, Urinetown |
| Special Citations: | Justin Bond and Kenny Mellman, Kiki and Herb: Jesus Wept; |
| Kirsten Childs, music and lyrics, The Bubbly Black Girl Sheds Her Chameleon Skin; |
| Rinde Eckert, And God Created Great Whales; |
| Mark Hollmann and Greg Kotis, book and lyrics, Urinetown; |
| Ed Kleban, music and lyrics, A Class Act; |
| Bob McGrath, director; and Cynthia Hopkins, Pilar Limosner, Bill Morrison, Laurie Olinder, Ruth Pongstaphone, Tim Schellenbaum, Howard S. Thies, Matthew Tierney, Fred Tietz, and Julia Wolfe, designers, Jennie Richee; |
| Ruben Santiago-Hudson; Lackawanna Blues |
| Sustained Achievement: | Marian Seldes |
| Ross Wetzsteon Grant: | Classical Theater of Harlem; |
| Clubbed Thumb; |
| Mint Theater Company; |
| Soho Rep; |
| Theatre for a New Audience |
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