Totti was born in Rome to parents Lorenzo and Fiorella Totti.

Although his youth club initially had come to an agreement to sell Totti to the Lazio youth side, one of Roma's youth coaches, Gildo Giannini, persuaded his parents to let him join the Roma youth squad in 1989.

After three years in the youth team, Totti made his first appearance for Roma's senior side in Serie A at the age of 16, when coach Vujadin Boškov called him up in the 2–0 away victory against Brescia on 28 March 1993.

Scored his first goal on 4 September 1994 in a 1–1 draw against Foggia.

Totti struggled to gain playing time under new manager Carlos Bianchi, and was even set to leave for Sampdoria on loan in January 1997. Roma's chairman at the time however, Franco Sensi, impeded the transfer, leading to further tensions with the manager, who eventually parted ways with the club.

Totti finished the league season reaching double figures in goals scored for the first time in his career; including a goal in the club's 2–0 away win against Napoli on 22 February 1998, Totti scored 13 goals in Serie A in total, and contributed to Roma's fourth-placed league finish that year.

Totti earned his first senior cap for Italy under the recently appointed international manager Dino Zoff during a 2–0 Euro 2000 qualifying victory against Switzerland on 10 October 1998.

31 October 1998, Totti became the official team captain, and the youngest Serie A club captain ever at the age of 22, inheriting the armband from Aldair.

In June 1999, Roma's then-president Franco Sensi sacked Zeman and signed Fabio Capello as a replacement, with the aim of winning the league title.

Totti was named the Serie A Italian Footballer of the Year for both 2000 and 2001.

Totti won the Serie A Footballer of the Year Award for the first time in his career in 2000.

Totti scored his first goal for Italy on 26 April 2000 in a 2–0 friendly win over Portugal in Reggio Calabria.

On 19 August 2001 Totti won his first Supercoppa Italiana, scoring and assisting in the 3–0 win against Fiorentina.

He scored his first hat-trick in Serie A, in a 3–2 away win over Brescia on 30 September 2002.

Totti also scored the only goal in Roma's decisive 1–0 victory at the Santiago Bernabéu Stadium against Real Madrid on 30 October 2002; this was the first time in 35 years that an Italian team had won a match in Madrid.

In 2003, Totti won his second Serie A Footballer of the Year Award.

Totti maintained consistent form by scoring 12 goals in Serie A, and 15 in all competitions, among which was his 100th Serie A goal from a free kick in a 3–3 draw against Internazionale, on 3 October 2004.

He scored his first brace for Italy in a 2006 FIFA World Cup qualification match in a 4–3 home win over Belarus, on 13 October 2004.

On 19 December, he became Roma's all-time leading scorer in Serie A after netting his 107th career league goal against Parma, breaking the record previously held by Roberto Pruzzo.

On 19 February 2006, he suffered a fracture of his left fibula and ligament damage during a match against Empoli after being fouled by Richard Vanigli.

Totti was at risk of missing the 2006 World Cup, but returned to the side on 11 May 2006 as a substitute in Roma's 3–1 Coppa Italia final defeat to Inter.

Italy won the World Cup 2006

Roma won the Coppa Italia final against league champions Inter with a 6–2 victory at home (Totti scored the opening goal after 50 seconds) and a 2–1 defeat at the San Siro in Milan.

On 17 June, he also received the ESM European Golden Shoe award as the top European goalscorer of the season.

Totti intended to retire from international football after the 2006 World Cup, but reneged on his decision and remained undecided on his future for over a year, not being called up in the meantime. He made his retirement official on 20 July 2007, at the beginning of the 2007–08 Serie A season, due to recurring physical problems and in order to focus solely on club play with Roma.

On 19 August 2007, Totti contributed to Roma's 1–0 victory over Inter in the 2007 Supercoppa Italiana, winning a penalty which was subsequently converted by Daniele De Rossi.

Totti scored his 200th goal for Roma in a 4–0 victory against Torino, in the Coppa Italia, on 16 January 2008.

On 9 March, he reached his 500th overall appearance for Roma, scoring the second goal of the match from a penalty kick in a 2–0 away win over Napoli.

Totti suffered a season-ending injury to his right knee during a 1–1 draw with Livorno on 19 April.

On 20 March, Totti became the sixth player in Serie A history to score 200 goals, finding the target twice in a 2–2 draw against Fiorentina to total 201 league goals.

On 31 May, in the final match-day of the 2014–15 season, he scored his 299th goal for Roma in a 2–1 home defeat to Palermo. Totti finished the season as Roma's leading scorer with ten goals in all competitions, the seventh time in which he had been the team's top scorer.

On 20 October, Totti made his 100th appearance in UEFA club competitions, setting up two goals in a 3–3 home draw against Austria Wien in the Europa League.

On 28 May, Totti made his final appearance for Roma in a 3–2 home win against Genoa, coming on as a substitute for Mohamed Salah in the 54th minute and received a standing ovation from the fans.

On 17 July 2017, Totti confirmed his retirement from professional football and announced that he had accepted an offer from his former team Roma to become a club director.

On 18 September, Totti began studying for a course in order to obtain his UEFA B coaching licence. However, he withdrew from the course two weeks later, as he was unable to attend the course regularly due to his commitments as a director of Roma; his former teammate Simone Perrotta took his place.

On 17 June 2019, he announced his resignations from his managerial role at AS Roma, in a hour-long press conference held at the Italian Olympic Committee headquarters where he publicly accused chairman James Pallotta and the management for not involving him enough into the club's decision making, up to the point of stating he would reconsider a comeback only under a different ownership. The events marked the end for his 30-year long formal association with the club.