"Mayo Chiki!" is a romantic comedy anime centered around Kinjirō Sakamachi, a high school student with a severe gynophobia—an intense fear of women. His condition stems from years of physical abuse by his younger sister, Kureha, and his mother, who is a professional wrestler. Kinjirō's life takes an unexpected turn when he discovers that Subaru Konoe, the popular and seemingly perfect butler serving the school's idol, Kanade Suzutsuki, is actually a girl in disguise. Subaru has been masquerading as a male butler to continue working for Kanade, as her family tradition dictates that only males can serve as butlers.

Kanade, aware of Subaru's secret, blackmails Kinjirō into keeping quiet by threatening to reveal his gynophobia to the entire school. In exchange for his silence, Kanade offers to help Kinjirō overcome his fear of women. This leads to a series of comedic and often awkward situations as Kinjirō interacts with Subaru, Kanade, and other female characters, including his childhood friend, Usami, who harbors feelings for him.

As the story progresses, Kinjirō's relationships with Subaru and Kanade deepen. Subaru, who initially resents Kinjirō for discovering her secret, gradually warms up to him, and a romantic tension develops between them. Kanade, on the other hand, enjoys manipulating Kinjirō for her amusement but also shows a softer side, hinting at her own feelings for him. The series explores themes of identity, gender roles, and personal growth, as the characters navigate their complex emotions and relationships.

The anime balances humor with moments of genuine emotional depth, particularly in its portrayal of Kinjirō's struggle to overcome his phobia and Subaru's internal conflict between her duty as a butler and her desire to live authentically. The interactions between the main characters drive the narrative, with each episode contributing to their development and the evolving dynamics of their relationships. "Mayo Chiki!" concludes with an open-ended resolution, leaving room for further exploration of the characters' futures.