Forrest is one of the 2nd-generation child units the player can recruit to their army once his father, Leo, has an S-rank with another female unit in your army. According to in-game support dialogue (pasted below), Forrest identifies as a heterosexual cisgender man, while not conforming to traditional gender roles/norms. Fans however have queerly read him to be trans or queer in other ways.
Corrin: I’ve never met anyone like you, and I’m curious: Do you prefer boys or girls?
Forrest: *giggle* Romantically speaking you mean? Girls, exclusively. I can’t imagine why you would have been confused on that point.
Forrest: Corrin … do you think I’m strange? Because of my interests and clothing that is. It’s not that I consider myself to be a girl. Far from it. But I see darling things such as this bracelet, and my heart simply melts. I wear what I feel most comfortable in, but I know your upbringing was … sheltered.
Moira O’Deorain is one of Overwatch’s playable characters. She is a geneticist, written as a villainous healer to balance out the number of heroic healers. When Overwatch was active, she was a member of its covert ops division, Blackwatch. After Overwatch’s disbandment, she was hired as Minister of Genetics in the city of Oasis. However, much of her work is actually funded by the terrorist organization, Talon, where she has a seat at the inner circle. Moira was conceived as an androgynous character, and is shown in her origin video wearing men’s clothing under her lab coat. Two of her cosmetic skins in the game alter her appearance to look like David Bowie to further emphasize these features.
The second great human country, Ydor, is the kingdom of art. Much more open minded than its neighbor Sarona, you can see openly gay couples, statues of naked men, and drag queens.
There is a steam achievement when playing this game called, “This is my hair, I don’t wear wigs,” that you can unlock if you speak to the drag queen wearing the wig hat, which is a reference to an Alaska Thunderfuck song.
Monokuma is a stuffed robotic bear, the self-proclaimed Headmaster of Hope’s Peak Academy, and the instigator of the Killing Game. In Chapter 5 of the game, there is a scene where Monokuma says the quote, “But in the bear kingdom, there IS no male or female!” but Makoto quickly turns Monokuma down by saying that there are male and female bears (found here).
Chihiro Fujisaki is one of the teens trapped inside Hope’s Peak Academy and forced to play the Killing Game. In the second chapter of the game, it is revealed that Chihiro is biologically male and dresses as a girl to deal with an inferiority complex. Chihiro was bullied as a child and told that he was weak for a man, resulting in him deciding to dress up as a girl as a way to be socially accepted as “weak”.
Many fans have speculated whether Chihiro is a trans character or not, but canonically, it seems that Chihiro identifies with the gender he was assigned at birth according to a conversation Chihiro has with Makoto during “School Mode” (found here) During the second chapter of the game, there are trans shock “she was a he?!” tropes played up as the rest of the cast find out about Chihiro’s secret (found here).