Tiny Toons Looniversity offers a positive and inclusive portrayal of LGBTQ+ characters and themes. It features explicit LGBTQ+ elements like drag as empowerment and openly gay characters such as Renaldo Raccoon, integrated naturally without stigma. The show promotes acceptance and understanding, depicting LGBTQ+ identities as a normal and affirming part of its diverse university world.

Tiny Toons Looniversity subtly addresses transsexual themes through positive portrayals of gender expression and drag culture. While not featuring explicit transsexual characters, the show promotes acceptance and diversity by challenging traditional gender norms in a playful and affirming manner, contributing to a supportive overall message.