The killers are homicidal murderers and recurring enemies that play a crucial role in Puppet Combo games, with the only exceptions of some games like Spiders, The Glass Staircase, and Samhain. Popular 70-80s horror film slashers (e.g: Michael Myers and Jason Voorhees) are the main inspirations for Puppet Combo's killers. Throughout the games, most killers serve as major threats in which they will pursue the protagonists and attempt to kill them, while a bit of few others take roles as general "protagonists" whom the player plays as.
- Main article: Neokalus Burr
- Main article: Driller Killer
The Driller Killer (real name unknown) is the main antagonist of Power Drill Massacre and a minor antagonist of Night Watch. He is a tall masked killer with a 3-year-old child's mind, and uses his deadly driller to slaughter his victims.
He originally appeared in the game's early incarnation Minotaur.
- Main article: The Mutilator
The Mutilator (real name unknown) is a crazed, dog-masked killer and the main antagonist of Meat Cleaver Mutilator.
- Main article: Night Ripper
The Night Ripper (real name unknown) is the titular main antagonist of the game of the same name. Only at nighttime, this duck-masked killer proceeds to chase and kill anyone whoever appear in his sight by stabbing their eyes with his pocket dagger.
- Main article: Santa
Santa is the main antagonist of all three story parts of Planet of Bloodthirsty Santa. He is a corrupted Santa Claus who was suborned into a blood-lusting murderer by some mysterious force, driving him to slaughter the whole elf village.
- Main article: Nun
The Nun (real name Janie mc.donnell) is the main antagonist of Nun Massacre where she haunts an abandoned Catholic school.
- Main article: Billy
He also appeared in Evil Clowns as one of the several enemies.
- Man article: Larry
The Riverside Ripper
- Main article: Unidentified Suspect
The Unidentified Suspect (real name unknown) is an unseen antagonist of The Riverside Incident, responsible for many killings, abducting, and setting houses on fire.
Night Shift Abductor
- Main article: Night Shift Abductor
- Main article: Easter Ripper
The Easter Ripper (real name Tom Smith) is the main antagonist of Murder House.