Ratonhnhaké:ton (1756 – unknown), also known as Connor, was a half English-half Kanien'kehá:ka Assassin who struggled to protect the Haudenosaunee as he fought for the Patriots during the American Revolutionary War. He developed and maintained a strong working relationship with George Washington, even after learning Washington's responsibility for his mother's death. He is an ancestor of Desmond Miles through the paternal line. Ratonhnhaké:ton is also the son of Haytham Kenway and the grandson of Edward Kenway. He is the main protagonist in Assassin's Creed III. He is voiced by actor Noah Watts.

Altaïr Ibn-La'Ahad (1165 – 1257) was a Syrian-born member of the Levantine Brotherhood of Assassins and served as their Mentor from 1191 until his death in 1257. During his tenure as Mentor, Altaïr made several discoveries and inventions that greatly helped the Order's progression. His leadership saw to the spread of the Assassins' influence in the Old World. With the Apple in hand, Altaïr changed the way members of his Order lived their lives, writing the details in his fabled Codex for later generations of the Order to read. Altaïr's vision of the Assassin Order was for them to be spread across the world, living among the people, and he began establishing many Assassins Guilds during his tenure as Mentor. Throughout his travels, Altaïr strengthened his Order, stopping various Templar plots over the years as well as halting the inexorable march of Genghis Khan.

Ezio Auditore (1459 – 1524) was a Florentine nobleman during the Renaissance and unbeknownst to most historians and philosophers, the Mentor of the Italian Brotherhood of Assassins, a title which he held from 1503 to 1513. He is also an ancestor to William and Desmond Miles. As well as Clay Kaczmarek. Ezio is the main protagonist of Assassin's Creed II,Assassin's Creed: Brotherhood, and shares the title of main protagonist with Altaïr Ibn-La'Ahad in Assassin's Creed: Revelations.He is also the main character of the short Assassin's Creed: Embers. He is voiced by actor Roger Craig Smith.

Edward James Kenway (1693 – 1735) was a Welsh privateer-turned-pirate, and an Assassin. He is the father of Haytham Kenway, the grandfather of Ratonhnhaké:ton, and an ancestor of Desmond Miles. Edward Kenway is the main protagonist of Assassin's Creed IV: Black Flag. He is voiced by actor by Matt Ryan

Shay Patrick Cormac (1731 – unknown) was an Irish-American Assassin-turned-Templar during the Seven Years' War. He is the main protagonist of Assassin's Creed Rogue. He is voiced by actor Steven Piovesan.