Tokyo Olympics: Wimbledon finalist Berrettini latest to join Games exodus
The world number eight, who had a bandaged leg in last week's Wimbledon final loss to Novak Djokovic, joins Rafael Nadal, Roger Federer and Stan Wawrinka in withdrawing from the men's event.
Serena Williams, Simona Halep, Victoria Azarenka, Johanna Konta, Bianca Andreescu and Angelique Kerber are among those to have already announced their decision to skip the women's tournament in Tokyo.
Berrettini, the first Italian to reach a Wimbledon singles final, announced his decision in an Instagram post on Sunday.
The 25-year-old's announcement comes a day after compatriot Francesco Molinari revealed he will not be taking part in the golf event at the Olympics.
"I am extremely disappointed to announce my withdrawal from the Tokyo Olympic Games," Berrettini said.
"I had an MRI scan yesterday on the thigh injury I sustained during Wimbledon and was informed I will not be able to compete for a couple of weeks."
The Olympic tennis events begin on July 24 and run through to August 1.
However, the deadline to name new athletes passed on Friday, so the Italian National Olympic Committee will not be able to name a replacement to join Fabio Fognini, Lorenzo Sonego and Lorenzo Musetti.
"Representing Italy is the biggest honour for me so it is devastating to miss the Olympics," Berrettini added.
"I wish the entire Italian team the best of luck in Tokyo. I will be supporting you all the way."
Despite the flurry of withdrawals, world number one Djokovic confirmed this week that he will enter the Olympic Games.
Djokovic travels to Japan in pursuit of a ground-breaking achievement, the Serbian just two titles – the Olympics and US Open – away from a first men's Golden Slam after triumphing at the Australian Open, French Open and Wimbledon in 2021