Williams' last US Open women's title came in 2014 against Caroline WozniackiSix-time champion Serena Williams has become the latest high-profile player to pull out of next week's US Open.
In a post on social media, the 39-year-old American great said she needed time to allow her body to "heal completely" from a torn hamstring.
Williams joins defending men's champion Dominic Thiem, plus Roger Federer and Rafael Nadal, in pulling out of this year's final Grand Slam.
The tournament, at Flushing Meadows in New York, begins on 30 August.
Williams wrote on Instagram: "After careful consideration and following the advice of my doctors and medical team, I have decided to withdraw from the US Open to allow my body to heal completely from a torn hamstring.
"New York is one of the most exciting cities in the world and one of my favourite places to play - I'll miss seeing the fans but will be cheering on from afar.
"Thank you for your continued love and support. I'll see you soon."
Williams suffered the injury to her hamstring during her first-round exit at Wimbledon in June, when the seven-time champion was forced to retire against Belarusian Aliaksandra Sasnovich.
The US Open presented Williams with another chance at a record-equalling 24th Grand Slam title.
She lifted her first major trophy at Flushing Meadows in 1999, defeating Swiss player Martina Hingis to become the first African American woman since Althea Gibson in 1958 - and the first in the Open Era - to win a Grand Slam honour.
After equalling Chris Evert's record of six titles in New York in 2014, Williams reached the final again four years later, but was beaten by Japanese rising star Naomi Osaka - a now three-time Grand Slam winner - in a controversial ending to the women's tournament.
In the men's event, Austrian Thiem has withdrawn due to a wrist injury, 20-time Grand Slam winner Federer is preparing to have further knee surgery, while world number five Nadal is out due to an injury to his left foot.
