Assassin's Creed: Revelations gained many of its concepts from a cancelled exclusive Nintendo 3DS title, Assassin's Creed: Lost Legacy. The game was announced at Nintendo's E3 2010 press conference in June, and was cancelled three months later, with Ubisoft's chief financial officer Alain Martinez stating that Ubisoft felt the console had enough "hardcore" games. The story of the cancelled title revolved around Ezio visiting Masyaf. Lead writer Darby McDevitt stated that the idea "morphed" into Revelations.

The absence of a missing right finger for the Levantine Assassins in Assassin's Creed was fixed in Revelations.

The original image along with the game's boxart shows that the Hookblade was originally going to be an upgrade for the left Hidden Blade, but in the final version of the game it was changed to replace Ezio's broken right Hidden Blade.

Although a crossbow weapon was removed from the first Assassin's Creed title due to being historically inaccurate, one still appears in one of Altair's memories, with Haras using it to execute a number of Assassins.

In the mission, "A Little Errand", if the player presses decline when the start-up appears, a glitch in the game will cause Ezio's hood to be lowered.

Sofia, Ezio's love interest in the game, was based on the Albrecht Dürer painting "Portrait of a Young Venetian Woman," which Art Director Raphael Lacoste was very adamant about using as the basis of the character. The painting can be seen in the Memory "Portrait of a Lady," where you have to retrieve it.