His decision follows fellow Australian Bernard Tomic making himself unavailable for Games selection last month after widespread criticism about his recent on-court attitude and behaviour.
"Unfortunately, while I have expressed every intention of trying to win a medal for my country in Rio, it's very clear to me that the Australian Olympic Committee has other plans," the world number 19 said in a statement.
"AOC's unfair and unjust treatment of me over the last four weeks, as well as the organisation's crystal clear position on whether they want me to be a part of the Australian Olympic team, has solidified my final decision."
She acknowledged Kyrgios' decision in a brief statement Friday, adding that "in regard to selection every athlete in contention is treated equally and fairly".
More From This Section
"We have no further comment on this issue."
Volatile 21-year-old Kyrgios is no stranger to controversy, frequently falling foul of tennis authorities.
Most recently, he received a code violation at the French Open for shouting at a ball boy and was fined for an audible obscenity during his thrashing at the hands of Richard Gasquet.
"While I have received assurances from Tennis Australia that I will be nominated for the Olympic team, the AOC has chosen to publicly and privately disparage me," he said.
"Not one member of the AOC has reached out to me, my family, my team, or representatives of Tennis Australia, asking for a meeting or the opportunity to discuss their concerns.
"The AOC's unwarranted attacks on me demonstrate the organisation's inability to understand the circumstances surrounding highly competitive sports."
TA president Steve Healy said Friday that Kyrgios continued to have the organisation's support.
"We understand Nick's decision and totally support him and his right to make it," Healy said. "But we are very disappointed that he has been put in this position.
"Nick is a passionate competitor and he's working hard to learn and mature in a highly pressurised environment where he is under constant public scrutiny."
Tennis Australia is due to make its Olympic selections on June 30, based on rankings. With Kyrgios and Tomic out of the running, John Millman (61) and Jordan Thompson (94) are next in line.