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A girl called Drew Decker (Carmen Electra) is home alone when she receives a phone call from an anonymous male caller who reveals that he is watching her. He then appears in a mask and costume, and attacks her with a knife. He stabs her in the chest, before she is then accidentally hit by her father's car as he returns home. The killer then finishes her off.

Cindy Campbell (Anna Faris) is at home as her boyfriend Bobby Prinze (Jon Abrahams) shows up. They have not yet consumated their relationship, but Bobby leaves without any change to this status.

The next day in school, Cindy meets with Bobby and the rest of her friends, Brenda Meeks (Regina Hall), Greg Phillipe (Lochlyn Munro), Ray Wilkins (Shawn Wayans), and Buffy Gilmore (Shannon Elizabeth). At the school, news teams are trying to interview the police and students who knew Drew. The most prominent among the reporters is Gail Hailstorm (Cheri Oteri), author of You're Dead, I'm Rich. Cindy points out to the others that Drew's murder happened exactly one year after they accidentally killed a man during a night of drunk driving, and dumped his body off a pier. Greg swears the group to a pact that they will reveal what happened to no one, and becomes physically abusive to Cindy when she exhibits reluctance.

During one of her classes, Cindy receives a note from someone telling her that they know about the incident with the man.

Buffy, competing in a beauty pageant, does a dramatic acting piece during the talent portion. During her reading, Greg is murdered by the killer in the balcony. Seeing this, Buffy attempts to warn everyone, but her desperate pleas are taken by the audience and judges as an exceptional acting performance, resulting in her winning the pageant.

That night, Cindy is attacked by the killer, but she calls the cops and the killer escapes. Bobby shows up with the same gloves and knife as the killer, and is arrested. Cindy goes to stay with Buffy that night but receives a call from the real killer, which results in Bobby's release. Bobby and Cindy later bump into each other at school, but their reconciliation is awkward, stemming in part from Cindy not getting over Ginger Spice leaving the Spice Girls.

The killer confronts Buffy in the girls' locker rooms. Making a sarcastic, metafictional observation about horror movies clichés, Buffy mocks the killer, even as he stabs her, and continues to do so even after he decapitates her.

Ray and Brenda go to the movies. Ray, while in a bathroom stall, peeps in on another occupant, and is impaled through the head with a penis. The killer then attempts to kill Brenda in the auditorium, but because of her uncourteous behavior, the other patrons kill her before he has a chance.

During a party at her house, Cindy has sex with Bobby. At the same time, the killer shows up at Shorty's house to kill him and his friends, but gets high on marijuana with them before doing so.

At Cindy's house, the killer shows up and apparently stabs Bobby. As Cindy attempts to attend to Bobby's wound, bobby shoots Shorty in the lung, causing marijuana smoke to exude. He reveals that the blood from his "wound" is ketchup, as he was never stabbed at all. Ray then appears, very much alive, and reveals that he and Bobby are the killer. When Cindy asks why they are doing this, they point out that they don't need one, because horror movies like Scream and I Know What You Did Last Summer do not boast logically coherent elements like plot or motive. Bobby also sarcastically suggests that it was lack of sex that drove him to do this, and reveals that he and Ray are gay lovers who will start a new life once they kill everyone who knows them, a crime they will get away with because they are merely copycatting a real serial killer who actually exists. To conceal their guilt, they decide to stab one another in order so that the authorities will believe them to be victims of the real killer. But after Ray stabs Bobby, the real killer appears and stabs Ray to death.

The killer then attacks Cindy, but after she repels him, he escapes. The police show up and take Cindy to the police station, where they find out that Buffy's brother Doofy (Dave Sheridan) was actually faking his mental retardation and is the real killer, and is in cahoots with Gail Hailstorm. After Doofy and Gail escape, the sheriff and Cindy find Doofy's disguise in the street and as Cindy cries out in the street over this discovery, she is hit by a passing car.

• Cindy Campbell - Played by Anna Faris, she is the parody character of both Sidney Prescott from the Scream trilogy and Julie James from the I Know What You Did Last Summer trilogy. Her name is a spoof of Sidney, and her surname is taken from Neve Campbell, the actress who played Sidney from Scream. Cindy is portrayed as naive and innocent compared to her friends. Her father is a drug dealer who works with the Escobar crime family ("Uncle Escobar," as Cindy knows him), dealing freebase and sometimes using it himself. To date, she is one of only two characters to appear in all of the Scary Movie films.
• Brenda Meeks - Played by Regina Hall, Brenda is Cindy's streetwise girlfriend. Brenda reappeared in Scary Movie 2, Scary Movie 3, and Scary Movie 4. She and Cindy are the only characters to appear in all of the Scary Movie films. She states that she doesn't know her dad in Scary Movie but in Scary Movie 3 Her parents are together and married.
• Bobby Prinze - Played by Jon Abrahams, he is the parody character of Billy Loomis from Scream and Ray Bronsen from I Know What You Did Last Summer and shares many of the same interests and personality traits with Billy, such as pressuring Cindy to lose her virginity to him. His name is taken from Freddie Prinze Jr., who starred in I Know What You Did Last Summer.
• Shorty Meeks - Played by Marlon Wayans, Shorty is the parody character of Randy Meeks from the Scream trilogy and Max from I Know What You Did Last Summer. He is Brenda's brother, and is depicted as a habitual marijuana user. He is later reappears Scary Movie 2.
• Ray Wilkins - Played by Shawn Wayans, Ray is the parody character of Phil Stevens from Scream 2. He also parodies Stu Macher from Scream towards the end of the movie. Although he is Brenda's boyfriend, numerous allusions appear in the film that he is homosexual, though he denies it. At the end of the film, he is revealed to be Bobby's accomplice. He is killed when the real killer (Doofy) stabs him in the back, though he is revived for the sequel. His name was taken from Ray Bronson, a character from the I Know What You Did Last Summer trilogy.
• Buffy Gilmore - Played by Shannon Elizabeth, Buffy is the parody character of both Tatum Riley from Scream and Helen from I Know What You Did Last Summer. Her name is a homage to Buffy the Vampire Slayer (as she parodies Helen Shivers, who is played by Sarah Michelle Gellar, who starred as Buffy the Vampire Slayer).
• Greg Phillipe - Played by Lochlyn Munro, Greg is the parody character of Barry Cox from I Know What You Did Last Summer. Greg has the same basic personality as Barry. His name is taken from Ryan Phillippe (who played Barry). Greg has an extremely small penis, and tends to be bellicose at times, and abusive towards Cindy for her desire to reveal their crime to the police. He is killed by Doofy at the pageant, parodying Ryan Phillipe's death in I Know What You Did Last Summer.
• Drew Decker - Played by Carmen Electra, Drew is the parody character of Casey Becker from Scream. Her name is taken from the actress who played Casey, Drew Barrymore. Her favorite horror movie is Kazaam. When the killer claims that isn't a horror movie, Drew simply tells him, "Well, you haven't seen Shaq act."
• Special Officer Doofy Gilmore - Played by Dave Sheridan, Doofy is the parody character of Dwight "Dewey" Riley from the Scream trilogy. Doofy is short for Doofus, as he suffers from mental retardation. As Buffy's 25-year-old brother, he still attends special education at B.A. Corpse High School. Doofy is a "Special Police Officer" whom nobody takes seriously. He enjoys masturbating with his mom's vacuum cleaner. At the end of the film, he is revealed to be one of the killers, and in a nod to the film The Usual Suspects, he is seen changing his look and mannerisms (his retardation having been faked), and getting away in a car with his accomplice, Gail Hailstorm.
• Gail Hailstorm - Played by Cheri Oteri, Gail is the parody character of Gale Weathers from the Scream trilogy. Ultimately, Gail and her long-suffering cameraman, Kenny, are stalked by the killer in a sequence parodying The Blair Witch Project. She is revealed to be Doofy's accomplice at the end of the film.

In addition to parodying the Scream series and I Know What You Did Last Summer, a number of other pop cultural references are made:

• The Blair Witch Project - The scene in which Gail Hailstorm and her Jamaican cameraman are videotaping one of the killer's murders, then they get spotted and they try to escape, however, Gail's cameraman gets killed and Gail apologizes to the cameraman's family by videotaping her apology and leaving it behind so it can be found.
• The Usual Suspects - Doofy's character and situation is similar to that of Verbal Kint's, and the coffee mug Cindy drops in the police station says "Doofus Porcelain", a direct parody of the scene where Detective Kujan discovers his own mug was made by a company called "Kobayashi Porcelain". In the ending scene, when Doofy walks away from the police station getting some clothes off and showing his real appearance while he smokes - that's like the Ending Scene of The Usual Suspects
• The Shining - When Buffy is about to be killed the word "Redrum," is whispered. In The Shining, Danny Torrance writes this word reversed, and speaks it, while semi-possesed.
• Titanic and Amistad - When Brenda and Ray are at the movies, a film trailer is shown showing a man on a ship at sea resembling like Captain Edward John Smith of the Titanic, and features a character (Keenen Ivory Wayans) saying the infamous "I'm king of the world!" line, leading the viewer to assume that the film is related to Titanic. Instead, a slave master whips the man, who is revealed to be a slave, as the trailer is revealed to be for a sequel to the 1997 film Amistad.
• Thinner - In the opening scene, Carmen Electra's character is run over by a car. The driver (her father) gets distracted in a similar way as in the movie Thinner.
• The Matrix - The fight scene between Cindy and the killer utilizes "Bullet time" special effects.