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Rule of Rose is a survival-horror game, with a focus on the effects of psychological horror and the impact of childhood trauma. The main character is Jennifer, a meek and easily intimidated young woman who follows a mysterious boy into an abandoned orphanage.

After some exploration, Jennifer finds a bloody sack, before having water poured over her by a group of children and being shoved into a coffin. Once she wakes up, she finds herself tied up on an airship heading to an unknown location.

It soon becomes clear that alongside various strangely shaped enemies, the only other living beings onboard are a few adults and the enigmatic Red Crayon Aristocrats, who hold a mysterious sway over the grown-ups.

Due to her mild disposition and fear, Jennifer is manipulated into bringing select offerings to this club ran by young girls each month. Her only loyal companion is Brown - the dog she finds and saves within the depths of the airship.

As Jennifer gradually recovers fragments of her past, it soon becomes clear that some truly horrific events have happened to her and most of the other characters. That does not mean her present will be any easier to survive though.