A member of the Templar Order, Ivan held one of the Staves of Eden. The Staves were Pieces of Eden used by the First Civilization. As a Templar dictator and wielder of a Piece of Eden, Ivan was a top target of the Assassins.
The Assassin Brotherhood dispatched Bykovski, a Livonian Assassin, to kill Ivan. As Bykovski prepared to strike Ivan as he passed during a parade, the Tsar's son, Ivan Ivanovich, spotted Bykovski and stabbed him repeatedly with a knife.
In 1581, Kirill, an Assassin and friend of Bykovski, infiltrated the Tsar's palace, intending to kill both Ivan and his son, the latter out of vengeance. After encountering Ivan Ivanovich, the Assassin stabbed the young heir in his left temple with the hidden blade. Kirill failed, however, to kill Ivan after becoming overwhelmed by guards and was forced to flee. Upon discovering his son's corpse, the hardened tyrant fell to his knees and wept while cradling his body.
Kirill successfully killed Ivan in 1583, after poisoning his drink as he played chess with an associate. In the confusion that followed, the Assassin recovered the Staff.
- The Templars would later manipulate history to claim that Ivan had murdered his son by beating him to death with the Staff following a heated argument.
- A painting by Ilya Repin of Ivan cradling his son's body was later used by Clay Kaczmarek as part of the glyphs. The glyph in particular, was covering Templars killed by the Assassins.
- The Brotherhood of Assassins would, ironically, use the Library of Ivan the Terrible as a repository.