The unreliable leader of "Yorozuya Gin-chan," a lazy sweet-tooth with silver permed hair and eyes like a dead fish. A self-indulgent man who loves Pachinko and Weekly Shonen Jump while falling behind on rent. However, his true identity is the legendary samurai once feared as the "White Demon" during the Joui War. Despite his usual lethargy, he shows overwhelming strength and chivalry when his friends or loved ones are in danger, heading to the battlefield with a single wooden sword to uphold his unyielding "Bushido (Soul)." He is a protagonist defined by the gap between his two sides.

A beautiful girl in a cheongsam with hair buns, but she is actually a survivor of the "Yato Clan," the strongest combat race in the universe, possessing superhuman strength. She is foul-mouthed, free-spirited, and gluttonous, often acting in ways unbefitting a heroine, but she is kind-hearted and lonely at her core. Her daily routine involves eating pickled seaweed and caring for the giant dog, Sadaharu. She ends her sentences with "-aru" and fights to protect her Yorozuya family while resisting her own violent combat instincts. The gap between her cute appearance, wild personality, and loyalty to her friends is her greatest charm.

The straight man of the Yorozuya. A serious boy working under Gintoki to revive his family's swordsmanship dojo. He is often teased for being plain and featureless, to the point where his "glasses" are called his true body. While he is a sensible and polite person, he relentlessly calls out the eccentricities of those around him. He has an otaku side as the fanatical captain of the Terakado Tsuu Imperial Guard. Though usually unreliable, he hides a samurai's pride and inner strength, bravely swinging his wooden sword when his friends are in trouble.

The Vice-Commander of the Shinsengumi. A cool and cold-blooded man feared as the "Demon Vice-Commander" for strictly enforcing the Kyokuchu Hatto (Station Rules). However, he is passionate and full of humanity at heart, with loyalty to the organization and the Commander that exceeds all others. He is an extreme "mayora" with a bizarre habit of piling mayonnaise onto every dish. He is also a heavy smoker. He and Gintoki are rivals who fight whenever they meet, yet respect each other's strength. A clumsy but dedicated man who follows his own Bushido, he is the Shinsengumi's number two.

The Captain of the Shinsengumi's First Division. A beautiful boy with refined looks, but inside he is a cold-blooded sadist known as the "Prince from the Planet Sadistic." He aims for the Vice-Commander position and tries to assassinate Hijikata with a bazooka whenever he sees an opening. However, his swordsmanship is the best in the Shinsengumi, and he ruthlessly cuts down enemies in battle. He is a habitual slacker who loves his eye mask. Though foul-mouthed and insolent, he deeply admires Commander Kondo and hides a hot-blooded soul that cares for his comrades and the Shinsengumi's pride.

The Commander of the Shinsengumi. A chivalrous leader who unites a group of rowdy soldiers through his personal virtue. He is often mocked as a "Gorilla" and has a playful side that leads him to repeatedly stalk Tae Shimura after falling in love at first sight. While he often serves as the comic relief involving dirty jokes or nudity, he is a pure and honest man at heart. He never abandons his comrades and is deeply respected by Hijikata and Okita like a father figure. He is the spiritual pillar of the Shinsengumi, unafraid to get his hands dirty and willing to risk his life to protect what matters.

Shinpachi's older sister. A reliable woman who supports the household by working at a snack bar to revive the family dojo. A beauty in a neat kimono, but her inner self is violent and terrifying; when angered, she possesses combat power that no one can oppose. She is especially ruthless in delivering iron-fisted punishment to Kondo for his stalking. She is a catastrophically bad cook, and every rolled omelet she makes turns into lethal "Dark Matter." However, she cares for her brother and the Yorozuya members more than anyone, and is a strong-willed woman with unwavering convictions.

Gintoki's childhood friend and the leader of a moderate faction of patriots seeking to overthrow the Shogunate. A handsome, genius swordsman known as the "Noble Fury," but he is extremely airheaded and prone to delusions. His catchphrase "It's not Zura, it's Katsura!" is well-known, and he uses various disguises to evade the wanted list. He dotes on his mysterious alien partner, Elizabeth. He shares a complex bond with his former classmates Gintoki and Takasugi, and while he spends his days being the butt of jokes, he hides a burning ambition for the future of the country.

Kotaro Katsura's partner, a mysterious alien creature. It looks like a person in a white penguin costume, with hairy human legs occasionally visible from underneath. It generally does not speak at all, communicating by writing on placards, though it occasionally drops biting truths or retorts. It shares a bond with Katsura that transcends words and is always by his side. Despite its surreal appearance, its combat ability is very high; it serves as a vital asset to Katsura's faction, even firing cannons from its mouth.

A kunoichi formerly of the Oniwabanshu, now a skilled assassin running a cleanup business. However, her true nature is that of an extreme masochistic stalker with an abnormal obsession for the protagonist, Gintoki Sakata. She is so nearsighted that she can see nothing without her glasses. She takes all of Gintoki's cold attitudes and insults as rewards, appearing unpredictably from ceilings or closets. Despite her perverted behavior, her combat skills as a kunoichi are top-tier, making her a reliable woman who will risk her life for her friends in a crisis.