| Julia Roberts | ... Charlotte the Spider (voice) |
| Dakota Fanning | ... Fern |
| Steve Buscemi | ... Templeton the Rat (voice) |
| John Cleese | ... Samuel the Sheep (voice) |
| Oprah Winfrey | ... Gussy the Goose (voice) |
| Cedric the Entertainer | ... Golly the Goose (voice) |
| Kathy Bates | ... Bitsy the Cow (voice) |
| Reba McEntire | ... Betsy the Cow (voice) |
| Robert Redford | ... Ike the Horse (voice) |
| Thomas Haden Church | ... Brooks the Crow (voice) |
| André 3000 | ... Elwyn the Crow (voice) |
| Dominic Scott Kay | ... Wilbur (voice) |
|
| Narrated by |
| Sam Shepard | ... Narrator (voice) |
|
| Kevin Anderson | ... Mr. Arable |
| Gary Basaraba | ... Homer Zuckerman |
| Beau Bridges | ... Dr. Dorian |
| Essie Davis | ... Mrs. Arable |
| Siobhan Fallon Hogan | ... Mrs. Zuckerman |
| Nate Mooney | ... Lurvy |
|
| Abraham Benrubi | ... Uncle the Pig (voice) |
| Louis Corbett | ... Avery |
| Julian O'Donnell | ... Henry Fussy |
| Robyn Arthur | ... Teacher |
| Nicholas Bell | ... Minister |
| Teague Rook | ... Well Dressed Man |
| Julia Zemiro | ... Bystander |
| Denise Kirby | ... New Teacher |
| Robert Plazek | ... Ball Thrower |
| Joseph Lotesto | ... Young Boy |
| Michael Roland | ... Mayor |
| Don Bridges | ... Announcer |
| Ian Watkin | ... Fair Official |
|
| Joel McCrary | ... Sheep Group (voice) |
| Brian Stepanek | ... Sheep Group (voice) |
| Fred Tatasciore | ... Sheep Group (voice) |
| Bradley White | ... Sheep Group (voice) |
| Maia Kirkpatrick | ... Joy the Baby Spider (voice) |
| Jennessa Rose | ... Aranea the Baby Spider (voice) |
| Briana Hodge | ... Nellie the Baby Spider (voice) |
|
| Puppeteers |
| Christopher Swift | ... Puppeteer |
| Matthew Heimlich | ... Puppeteer |
| Peter Chevako | ... Puppeteer |
| Patrick Rummans | ... Puppeteer |
| Philip Millar | ... Puppeteer |
| Peter Luscombe | ... Puppeteer |
| Kathleen MacGowan | ... Puppeteer |
| Michael Orlnelaz | ... Puppeteer |
| Malcolm J. Martin | ... Puppeteer |
