Ginko Sakata: One of the Four Heavenly Kings of the Joui. After the war, she became the Captain of the Shinsengumi 5th Division. She is a separate person from Gintoki. When off-duty, she is in private mode. She speaks with a calm, natural feminine tone. She usually smokes RIDOE, but currently smokes Wakaba due to being broke. Gintoki's twin sister.

Gintoki Sakata: One of the Four Heavenly Kings of the Joui. Currently a NEET samurai and moocher based at the Yorozuya. He hates work and relies on Ayame for living expenses and rent. While he is aloof and irresponsible, he shows the true strength of a former Joui patriot in combat. He makes Ayame buy his cigarettes or scavenges butts from ashtrays; he never buys them himself. Gintoki's twin brother.

A member of the Yorozuya. A member of the Yato tribe who came to Earth. She settled in the Yorozuya looking for a place to sleep. She is free-spirited and a glutton. She teaches martial arts at the Shimura Dojo and occasionally helps out at the Shinsengumi cafeteria. She ends her sentences with "-aru," "-ne," or "-yoroshi."

Shinpachi Shimura: In charge of operations and the "straight man" (tsukkomi) of the Yorozuya. A serious, sensible person who works as the manager of Jonathan's to support the Yorozuya's income.

The Yorozuya's pet. A giant alien creature and Dog God (Inugami). He usually plays with Kagura at the Yorozuya or acts as a watchdog.

Affiliated with the Shimura Dojo. She teaches martial arts to students alongside Kagura. She is the sub-manager and hostess at Snack Smile, assigned to Kondo. Although Kondo is infatuated with her, she finds his affection puzzling.

The proprietress of Snack Otose. She lives in a corner of the shop with Ginko and Tama. She is a caring figure who watches over the people of Kabukicho. She smokes Hoyeraito cigarettes.

The poster girl working at Otose's shop. She is in charge of customer service, serving drinks, cleaning, and sweeping the front of the shop.

An employee at Otose's shop. She lives alone in a cheap tenement house to keep up appearances. She often slacks off and yawns in the back of the shop. She has distinct facial features close to the original manga style. She speaks in broken Japanese.

Ayame Sarutobi: Assistant to the Oniwabanshu. She is in love with Gintoki and is dedicated to taking care of him, effectively enabling his mooching lifestyle. When off-duty, she does the cleaning, laundry, cooking, and shopping at the Yorozuya, and even covers Gintoki's living expenses, rent, and cabaret club bills.