Jasmine Paolini defeated Coco Gauff in the final, 6–4, 6–2 to win the women's singles tennis title at the 2025 Italian Open. It was her second WTA 1000 and third career WTA Tour singles title. Paolini was the second Italian woman in the Open Era to win the Italian Open singles title, after Raffaella Reggi in 1985. The final was also a rematch of the doubles final from the previous year. Paolini was the first Italian woman to win both the singles and doubles titles at the tournament in the same year since Reggi in 1985, and the first woman to do so overall since Monica Seles in 1990.
Iga Świątek was the defending champion, but lost in the third round to Danielle Collins.
Gauff was the seventh and youngest woman to reach the finals of both Madrid and Rome in the same year. Peyton Stearns became the first player in the Open Era to win three straight WTA Tour matches in third-set tiebreakers, against Madison Keys, Naomi Osaka, and Elina Svitolina in the third round, fourth round, and quarterfinals respectively.
Seeds.
All seeds received a bye into the second round. 
Draw.
Finals.
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Seeded players.
The following are the seeded players. Seedings are based on WTA rankings as of 21 April 2025. Rankings and points before are as of 5 May 2025.
Under the 2025 Rulebook, points from six of the seven combined WTA 1000 tournaments (which include Rome) are required to be counted in a player's ranking.
† The player withdrew from the 2024 tournament but was not required to carry a 0-point penalty in her ranking due to a long-term injury exemption. Points from her 18th best result will be deducted instead.
Withdrawn seeded players.
The following player would have been seeded, but withdrew before the tournament began.
Other entry information.
Withdrawals.
‡ – withdrew from entry list
§ – withdrew from main draw