The film is set approximately one year after the "requel" events of Scream 5 in Woodsboro. The four survivors sisters Sam Carpenter and Tara Carpenter, along with twins Mindy Meeks-Martin and Chad Meeks-Martin have relocated from California to New York City in an attempt to start a new, quiet life and escape the trauma of Ghostface.

Sam continues to struggle with the revelation that she is the daughter of the original Ghostface killer, Billy Loomis, and attends therapy to manage her mental state, often fearing she may inherit her father’s violent tendencies. Tara is attempting to live a normal college life, but their new environment is immediately complicated by the massive, anonymous population of the metropolis.

The new wave of Ghostface attacks begins with a brutal murder in a crowded subway car. This Ghostface is more aggressive, cunning, and resourceful than previous iterations, utilizing the anonymity and chaos of NYC to their advantage. They leave behind masks from past killers at each scene, indicating a deep obsession with the entire lineage of the Woodsboro murders.

Sam and Tara quickly realize their attempt at a fresh start has failed. They are again the targets, and the high-profile nature of the NYC murders draws immediate national attention. They are forced to rely on their inner circle, which includes Gale Weathers, who flies in to cover the story, and Kirby Reed, the lone survivor of Scream 4, now an FBI agent who specializes in Ghostface cases.

The core mystery revolves around Ghostface’s identity and their motive, which seems to be driven by a desire to complete the work of the previous killers or to punish Sam for being Billy Loomis's daughter. Suspicion falls upon several new characters in their college circle, including Ethan Landry, Quinn Bailey, and Detective Wayne Bailey, a police officer and Quinn's father.

The climax takes place in an abandoned, decaying movie theater that has been converted into a macabre shrine dedicated to the Ghostface killers, filled with costumes, weapons, and memorabilia from all the previous massacres. Sam, Tara, and the twins are lured there for the final confrontation.

In a dramatic reveal, it is exposed that there are three Ghostface killers, all from the same family: Detective Wayne Bailey, and his children, Ethan Landry and Quinn Bailey. Their motive is simple: they are the family of Richie Kirsch (one of the killers from Scream 5), seeking revenge on Sam and Tara for killing him.

The final, chaotic battle is a multi-front fight for survival within the shrine. Sam, embracing her darker, survivalist instincts (and briefly utilizing a Ghostface mask and knife in a chilling echo of her father), teams up with Tara to dispatch the vengeful family. Sam and Tara kill all three killers, ending the threat.

The film concludes with the Carpenter sisters choosing to stay in New York, facing their trauma head-on. Tara finally acknowledges Sam’s complicated legacy and accepts her as her sister, solidifying their bond, while the legacy of Ghostface remains, haunting the city and the survivors.