"Pani Poni Dash!" is a surreal and fast-paced comedy anime that revolves around the chaotic daily lives of students and faculty at an all-girls high school. The central figure is Rebecca Miyamoto, a child prodigy who becomes a teacher at the age of 11. Despite her young age, Rebecca, often called "Becky," is highly intelligent and confident, though her eccentric personality and unpredictable behavior frequently lead to absurd situations. Her class, consisting of a quirky group of students, includes the energetic and mischievous Himeko Katagiri, the stoic and deadpan Ichijo, the timid and easily frightened Rei Tachibana, and the mysterious transfer student Kurumi Momose, who often breaks the fourth wall and interacts with the audience.

The series lacks a conventional overarching plot, instead presenting a series of episodic vignettes filled with rapid-fire jokes, parodies of other anime and pop culture, and surreal humor. The narrative often shifts tone abruptly, blending slapstick comedy with moments of absurdity and meta-commentary. The school setting serves as a backdrop for a wide range of comedic scenarios, from mundane classroom antics to bizarre supernatural occurrences, such as encounters with aliens, ghosts, and anthropomorphic animals.

Supporting characters add to the chaotic atmosphere, including the school's principal, who is often seen wearing a rabbit costume, and the overly dramatic and melodramatic vice-principal. The series also features recurring gags and running jokes, such as the frequent appearance of a sentient, talking cat named Mesousa, who often provides sarcastic commentary on the events unfolding around her.

The humor in "Pani Poni Dash!" is heavily reliant on cultural references, wordplay, and visual gags, making it a dense and fast-paced viewing experience. The narrative structure is non-linear, with episodes often jumping between unrelated skits and scenarios, creating a sense of unpredictability and randomness. Despite its lack of a traditional storyline, the series maintains a cohesive tone through its consistent use of absurdity and its focus on the eccentric personalities of its characters. The interplay between Becky and her students, as well as the interactions among the students themselves, drives much of the humor and keeps the narrative engaging.