Orphaned from an early age, Paola had known many hardships during her life, until she was finally charged with the murder of a city guard. With the help of Giovanni Auditore da Firenze, Paola managed to get out of trouble, after which she founded her brothel.
After Giovanni's death at the hands of the Templars, Paola assisted the young Ezio Auditore da Firenze multiple times, teaching him how to blend and lower his notoriety. She also renewed his contact with Leonardo da Vinci, who was able to repair his Hidden Blade.
Paola was born in 1438, as the daughter of two members of the Assassin Order. When she was eight years old, her parents were killed at sea and Paola and her sister Annetta were left without care. Due to this, when she was given the choice to join either a nunnery or a brothel, Paola chose to live a life on the streets instead.
After having survived on the streets for 20 years, she was arrested for murdering a city guard. However, Giovanni Auditore, a member of the Assassin Order, represented her in court and won the case on a self-defense plea.
After that, Paola became an Assassin herself and founded a brothel, which she named "La Rosa Colta", to help other women who had been forced to live on the streets. Annetta, meanwhile, became the Auditore family's live-in servant.
In 1476, Giovanni Auditore and two of his sons were executed by the Templars, under the false charge of treason against Florence. Annetta brought Giovanni's wife Maria and daughter Claudia to Paola to seek refuge, where they were soon joined by Giovanni's remaining son Ezio.
Soon after, Paola taught Ezio how to remain undetected by the city guards, as Ezio aspired to take revenge on Uberto Alberti – the man who had arranged his family's execution. Afterwards, she sent him to Leonardo da Vinci, who would be able to repair Giovanni's broken Hidden Blade.
After Ezio had the blade repaired and used it to kill Uberto, he returned to Paola, who informed him of ways to draw the city guard's suspicion away from him. On having done so, Ezio took his mother and sister to the safety of Monteriggioni.
In 1488, Paola and some of the other Assassins gathered in Venice to acquire the Apple of Eden, an ancient artifact which was to be brought to Venice by the Templars that day. With this aim, they ventured to the location where Rodrigo Borgia, Grand Master of the Italian Templars, would retrieve the Apple from his men.
Arriving there, the Assassins stumbled upon Ezio fighting off Rodrigo and his guards. While Paola, Teodora Contanto and Niccolò Machiavelli remained on the sidelines, the other Assassins assisted Ezio in the fight. At the fight's conclusion, Rodrigo fled and Ezio took the Apple, followed by Niccolò revealing to him that all of the others were Assassins.
In 1494, Girolamo Savonarola took control of Florence – which had been left without a strong leader after the death of Lorenzo de' Medici – with the help of the Apple that he had obtained from Ezio six years earlier.
In 1497, Savonarola instigated the Bonfire of the Vanities, where he forced all of the Florentines to destroy any form of progression into the Renaissance. Niccolò met with Paola, and informed him that Ezio was going to assassinate all of Savonarola's lieutenants and that they would need to lead the citizens into a revolt.
After all nine lieutenants had been killed, Paola, Ezio, La Volpe and Machiavelli witnessed a mob gathering in front of the Palazzo Pitti to take Savonarola captive. Shortly afterwards, Paola was also present at Savonarola's burning, though Ezio killed him before the fire reached him, and gave a speech about the people who composed the guiding force behind his actions, and that men have the privilege of free will.
In 1499, Paola traveled to the Villa Auditore to help formulate the plan to assassinate Rodrigo Borgia. She and the others distracted the city guard in Rome while Ezio infiltrated the Sistine Chapel.
After that, Paola kept contact with the Auditore family, especially Claudia, with whom she had become close friends. In 1512, Claudia mentioned Paola was still alive and a leader of the Assassins.