The game features 43 playable Assassins such as Ezio Auditore da Firenze, Aguilar de Nerha, Shao Jun, Claudia Auditore da Firenze, and Niccolò Machiavelli, along with more than 20 new unlockable characters exclusively created for Rebellion.
The game features a headquarters for the Brotherhood where Assassins can be trained and new items and equipment can be crafted. As the player levels up their brotherhood, additional rooms and other content become available, opening up new upgrade options.
The player can also send teams of three Assassins on various infiltration missions. Different skills and character combinations are better suited to some missions than others. In addition, the type of missions completed will have an effect on how the Assassins develop: standard missions are key for gathering resources to upgrade the player's Assassins, while Loot missions will earn materials for crafting.
New Assassins are unlocked by collecting their DNA fragments in Story or Legacy missions.