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Its director Dee Rees, from the United States, will now use the money to make another short film in the UK. Her film Pariah was chosen by judges as the best entry of the 30 shortlisted at the end of a three day film festival in Cardiff.
It also won the NewFest festival award in New York earlier this year.
Ms Rees' 27-minute film follows the teenage girl as she unsuccessful tries to juggle multiple identities to avoid rejection from her friends and family.
A coming-of-age drama about a lesbian teenager has won the £25,000 Iris Prize - thought to be the largest ever award for a gay and lesbian short film.
Its director Dee Rees, from the United States, will now use the money to make another short film in the UK.
Her film Pariah was chosen by judges as the best entry of the 30 shortlisted at the end of a three day film festival in Cardiff.
It also won the NewFest festival award in New York earlier this year.
Ms Rees' 27-minute film follows the teenage girl as she unsuccessful tries to juggle multiple identities to avoid rejection from her friends and family.
Announcing the winner of the prize at the awards show at Cardiff's Cineworld, Frances Hendron, jury chair said: "This film was impactful. It is a personal story told in a universal manner.
"The film maker displays excellent film making skills from the direction of the actors, the integral music sound track and a natural control of its subject."
The winner of the best British entry at the Iris Prize was Private Life by film-maker Abbe Robinson, who studied at Newport.
The film is set in Yorkshire in the 1950s and tells the story of Ruth Ackroyd who leaves her father's textile mill and begins leading a secret double life in Manchester.
The judges also commended four films: For the Love of God by British film-maker Joe Tucker, Peace Talk by Swedish director Jennifer Malmqvist, Hello, My Name is Herman by Canadian Karine Silverwoman and Le Weekend by Timothy Smith.
"Strong audiences"
Debates, workshops and screenings of gay and lesbian feature films were held at Chapter and Cineworld in Cardiff and organisers said the festival attracted "strong audiences".
Films ranged from the highly acclaimed The Bubble, which tackled same sex love across the Israeli-Palestinian divide, to spoof, camp horror movie Gay Bed and Breakfast of Terror.
Festival founder Berwyn Rowlands said plans were already underway for the Iris Prize Festival to make a return in 2008.
He added: "We've demonstrated during this inaugural festival that Iris has many friends located throughout the world.
"Filmmakers who joined Iris in Cardiff came from the Bahamas, New York, Los Angeles, Toronto, Berlin, Paris and London."
http://news.bbc.co.uk/2/hi/uk_news/wales/7032376.stm