The 32-year-old revealed through social media that he wouldn't be part of the Paris push following a selection camp in Darwin over the weekend.
The squad was finalised by coach John Manenti, with Hooper's hopes dashed by an injury-plagued transition from 15-a-side rugby.
View this post on Instagram A post shared by Michael Hooper (@mik.hooper)
"What a ride! My Olympic journey has come to an end, and with it, my Australian Rugby career," Hooper posted on Instagram.
"To the Aussie Sevens team, I wish you all the best you've been an awesome, helpful, and talented group of athletes to be part of.
"As for my time in Australian Rugby, I'm grateful to my family and friends for their unwavering support throughout. We've shared an incredible journey, filled with unforgettable experiences, challenges, and memories. Thank you."
Ending a legendary career, the former NSW Waratahs flanker played 125 Tests and captained the Wallabies more than any other player, leading the men in gold on 69 occasions.
He made his Test debut in 2012 as a 20-year-old and two years later became the youngest to captain Australia since Ken Catchpole, and also became the youngest to play 100 Tests.
Hooper won four John Eales Medals, a player-voted award for the Wallaby of the year, while he was twice nominated for the World Rugby Men's 15s Player of the Year.
Michael Hooper played 125 Tests and led the Wallabies a record 69 times through his storied career. (Jason O'BRIEN/AAP PHOTOS)
After controversially missing selection in the Wallabies' squad for the 2023 Rugby World Cup, Hooper switched to sevens late last in an attempt to end his career at the Olympics.
But a series of injuries delayed his debut until April and he was only able to play two world series tournaments before suffering a groin injury, which ruled him out of the final in Madrid.
Rugby Australia CEO Phil Waugh said Hooper had been incredible player and ambassador for the sport in Australia.
"He leaves a legacy that will remember him as one of the greatest to wear the Wallaby gold and a favourite among Australian supporters," Waugh said in a statement."Michael's incredibly well respected here and around the world, and on behalf of Rugby Australia, I want to wish him and his family all the best for the future."
The Australian sevens team will be announced by the AOC on Wednesday.
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