It is the second live-action feature-length film starring the Looney Tunes characters, the first being Space Jam (1996). It was theatrically released on November 14, 2003. The film was a box office bomb, grossing only $68.5 million against its $80 million budget.
Tired of playing second fiddle to Bugs Bunny, Daffy Duck demands his own movie from the Warner Bros., but they promptly fire him. Aspiring stuntman DJ Drake is asked to escort Daffy off the studio lot, but the ensuing chase leads to the Batmobile demolishing the studio water tower. DJ is fired, and returns home with Daffy hitching a ride, discovering his father and action star Damian Drake is actually a secret agent. Damian instructs his son to travel to him, find his associate Dusty Tails, and gain the mystical Blue Monkey diamond before being captured by the Acme Corporation, led by the childish Mr. Chairman. DJ and Daffy head for him. Meanwhile, Bugs' routines fall apart without Daffy, so V.P. for Comedy, Kate Houghton, is sent to rehire Daffy or face being fired herself. Bugs informs her of the situation and they pursue DJ and Daffy in Damian's spy car.
In DJ and Daffy meet Dusty in a casino owned by Acme operative Yosemite Sam. Dusty gives them a strange playing card, but when Yosemite tries to kill them, they flee in the spy car with Bugs and Kate, but when Daffy cries "Mother!", the car takes flight. Crashing in Death Valley, the group stumble across the secretive Area 52, run by Damian's gadget specialist Mother. Mother plays a video recording, revealing Acme intends on using the Blue Monkey to transform mankind into monkeys to manufacture their products and then buy them as humans. Marvin the Martian, imprisoned in the facility, escapes and leads a group of fellow alien inmates to obtain the playing card but the heroes escape. Realising the card has the Mona Lisa's face on it, the group conclude they must view the painting in the Louvre, Paris.
At the Louvre, the group discover the card contains a viewing window and looking through it reveals the Mona Lisa has a map of Africa hidden beneath it. Elmer Fudd appears, revealing he is "secretly evil", and chases Bugs and Daffy through the gallery for the card whilst Kate is kidnapped by Mr. Chairman's bodyguard Mr. Smith to obtain a photo of the African map though DJ rescues her. Elmer is disintegrated after jumping out of a pointillism artwork by Bugs.
DJ, Kate, Bugs, and Daffy travel to Africa, meeting Granny, Sylvester, and Tweety, who escort them to the ruins of a jungle temple where they find the Blue Monkey. Granny and co. reveal themselves to be Mr. Chairman, Smith, and the Tasmanian Devil in disguise. Mr. Chairman uses a disintegration gun to transport himself and the heroes to the Acme headquarters where he forces DJ to give him the diamond when Damian is revealed to be his prisoner.
Marvin is sent to place the Blue Monkey on an Acme satellite which will emit an energy beam around the world to turn everyone, save Mr. Chairman, into monkeys. DJ and Kate rescue Damian from a death trap, whilst Bugs and Daffy pursue Marvin. Bugs is incapacitated, prompting Daffy to become Duck Dodgers, removes his beak and uses it to destroy the Blue Monkey. The transforming energy beam only strikes Mr. Chairman, turning him into a monkey.
Bugs and Daffy return to Earth, where Daffy discovers the whole adventure was staged to make a movie. However, Bugs promises Daffy they will be equal partners, but just as Daffy's luck seems to be improving, he is flattened by the Looney Tunes' iris, where Porky Pig tries to close the film with "That's all folks!" only for the studio to shut down before he can finish and he tells the audience to go home.
- Joe Alaskey as Bugs Bunny, Daffy Duck, Sylvester and Tweety
- Brendan Fraser as D.J. Drake, Himself
- Jenna Elfman as Kate Houghton
- Steve Martin as Mr. Chairman
- Heather Locklear as Dusty Tails
- Joan Cusack as Mother
- Timothy Dalton as Damian Drake
- Bill Goldberg as Mr. Smith
- Don and Dan Stanton as Mr. Warner and Mr. Warner
- Matthew Lillard as Himself (Cameo)
- Jeff Gordon as Himself (Cameo)
- Kevin McCarthy as Dr Miles Bennell (cameo)
- Michael Jordan as Himself (Archive footage from Space Jam)
- Marc Lawrence as Acme VP, Stating the Obvious
- Ron Perlman as Acme VP, Never Learning
- Bob Bergen as Porky Pig
- June Foray as Granny and Mama Bear
- Billy West as Elmer Fudd, Papa Bear, Junior Bear, Nasty Canasta, Larry, Rancid Rabbit
- Bruce Lanoil as Pepé Le Pew
- Casey Kasem as Shaggy
- Frank Welker as Michigan J. Frog, Marvin the Martian, Speedy Gonzales, Beaky Buzzard, Scooby-Doo and The Roadrunner
- Jeff Bennett as Yosemite Sam and Foghorn Leghorn
- Caroll Spinney as Oscar the Grouch (Cameo) (Footage from Sesame Street)