Charon ferries damned souls across the River Acheron into Limbo, the First Circle of Hell. His physical form manifests as a colossal amalgamation of an anthropomorphic vessel and a giant ship integrated into his body, lacking conventional limbs beyond his head. This design evolved to accommodate the immense volume of souls arriving over millennia.

He strictly enforces the prohibition against living beings entering Hell, refusing passage to Dante upon their first meeting. When Dante demands information about Beatrice's location, Charon cryptically reveals Dante's life and soul already belong to Hell and mentions Beatrice made "a very foolish wager," alluding to her bet with Lucifer concerning Dante's faithfulness during the Crusades.

Unable to attack directly due to his physical constraints, Charon summons demonic minions to assault Dante for violating underworld laws. This confrontation ends with Dante using an Asterian Beast to decapitate Charon, killing him. Charon's severed head later appears within the Citadel of Limbo, endlessly repeating the inscription "Abandon all hope, ye who enter here" from the Gates of Hell. His disembodied head resurfaces in the River Styx, functioning as a nest for Pests.

His role and design draw inspiration from Greek mythology, where he ferries souls across the Acheron or Styx, requiring payment via coins placed on the deceased's tongue. While the film adaptation transforms his physical appearance into a living vessel, it retains his core mythological function as the boundary keeper between the living world and the damned.