At the Budokan, an annual event known as the Tsukino Concert puts Tsukino Entertainment Production's best and brightest idol units on the stage. The leaders of idol groups SolidS, SOARA, QUELL, and Growth must compose four unique and distinctive songs for their respective units, which they will perform on the second day of the festival.

Composing four new songs proves to be a difficult task for SolidS's leader, Shiki Takamura, as he struggles to find inspiration. Sora Oohara must learn to overcome past mistakes while propelling his unit, SOARA, to the top. Wanting to escape his past stardom, Shuu Izumi is determined to push his unit, QUELL, up to the level of popularity that his previous one had. Growth finds themselves working together with a rival group, ZIX, prior to their debut—something that leader Kouki Etou is reluctant to do.

Tsukipro The Animation tells the stories of these four units simultaneously as they go through their daily lives while preparing for the Tsukino Concert.

Tsukipro is based off of a multi-media franchise under the same name by Movic (credited as Tukino Talent Productions). It is a collaboration project between popular Japanese voice actors and Vocaloid producers. The franchise has spawned multiple music CDs and drama tracks, as well as a mobile rhythm game, entertainment app, manga, online radio shows, 2.5D dance live shows and a live action movie. Tsukipro was created following the success of Tsukiuta., another project under the same company.