Key & Peele occasionally features LGBTQ+ characters and themes within its sketch comedy. These portrayals are generally incidental, often using situational humor or satirizing societal reactions rather than directly affirming or denigrating queer identity. The humor typically arises from external circumstances or other characters' perspectives, resulting in a neutral overall impact.

Key & Peele includes a sketch where a criminal character falsely claims a trans identity to manipulate the legal system. This depiction uses gender identity as a source of mockery and a deceptive tactic for comedic effect. The portrayal reinforces harmful stereotypes by presenting trans identity as a tool for manipulation, leading to a net negative impact on the holistic representation of transsexual themes.