The International Olympic Committee (IOC) is banning transgender women from competing in all female events.
The restriction applies to eligibility for any female category event at the Olympic Games, or any other IOC event, including individual and team sports.
The policy is not retroactive, and does not apply "to any grassroots or recreational sports programmes", the IOC said in a statement, adding that it "protects fairness, safety and integrity in the female category".
Those events will now be limited to biological females, determined on the basis of a one-time test for the SRY gene, which "is fixed throughout life and represents highly accurate evidence that an athlete has experienced male sex development", the IOC said.
The organisation's new policy aligns with US President Donald Trump's executive order on women's sports ahead of the 2028 Los Angeles games, the first Olympics where the new rules will apply.
It is unclear how many, if any, transgender women are competing at Olympic level.
No woman who transitioned from being born male competed at the Paris 2024 Olympics.
Runner Caster Semenya, a two-time Olympic champion who has a medical condition known as differences in sex development (DSD), is among those affected by the change.
IOC President Kirsty Coventry established a review of "protecting the female category" soon after becoming the first woman to lead the Olympic body in its 132-year history last June.
In a U-turn to previous policy, she wanted to bring in a universal rule for competitors in female elite sports after years of fragmented regulation that led to some major controversies.
In the statement, she said that even the smallest margins "can be the difference between victory and defeat".
Ms Coventry added: "So, it is absolutely clear that it would not be fair for biological males to compete in the female category. In addition, in some sports it would simply not be safe."
Female eligibility was one of the main issues in the IOC election last year when Coventry's main rivals said they would adopt a stronger policy.
Prior to the games in Paris, track and field, swimming and cycling had already passed rules excluding transgender women who had been through male puberty.
Being born male gives physical advantages that are retained, the IOC said, pointing to its own research.
Read more on Sky News:
Trans girls' Guiding deadline
Puberty blocker trial paused
"Males experience three significant testosterone peaks: in utero, in mini-puberty of infancy and beginning in adolescent puberty through adulthood," the document said.
It added this gives males "individual sex-based performance advantages in sports and events that rely on strength, power and/or endurance."
Sky sports correspondent Rob Harris called it a "landmark decision" driven in part by "growing concerns about the potential for physical advantages that are retained by transgender women who have gone through male puberty".
The IOC sees itself as "protecting the female category at the Olympics for biological females", as well as ensuring competitors' safety, especially in sports like boxing, where physical advantages "could potentially be dangerous".
There was "big pressure" on the IOC, not least from Mr Trump, who, at a fundraiser on Wednesday, "was repeating this desire to keep transgender women out of women's sporting events", Harris said.
(c) Sky News 2026: Transgender women athletes banned from female Olympic events
Gang-rape victim, 25, to be euthanised after father's legal challenge fails
Sarah Ferguson stripped of 'freedom of York' by councillors over ties to Jeffrey Epstein
Police reopen investigation into Andrew Tate sex abuse allegations
UK facing biggest economic hit from Iran war of any major country
Greens pile pressure on Ed Miliband to end 'rigged energy system'
Nicolas Maduro returns to US court as lawyer fails to dismiss drug trafficking charges
Lebanon doctors accuse Israel of repeating Gaza's 'scorched earth policy' with attacks on hospitals and staff
Labourer who killed mother-of-two and buried her in shallow grave jailed for 18 years