The side-scrolling platformer features a two player co-operative mode over local wireless, which needs two game cards and two Nintendo 3DS systems, and it is not capable of online multiplayer.
The game is a port of the Wii game of the same title and includes an easier mode to "better optimize [the game] for portable playability." In the new mode, Diddy and Donkey can take up to five hits from an enemy thanks to the increase of hearts from two to three. The easier difficulty setting also includes extra environmental objects to assist Donkey and Diddy, such as balloons and crash guards.
The title also features an Original Mode which sets the difficulty to the level of the original Wii version.