American Dad! incorporates LGBTQ+ characters and themes, notably through Roger's fluid identity and diverse personas. While often used for comedic effect and sometimes employing stereotypes, these portrayals generally avoid endorsing degradation. Queer identity is present and contributes to character complexity and plot points without being the sole source of ridicule or affirmation, leading to a largely neutral overall impact.

American Dad! features a significant transsexual character arc in the episode "Tuttle's Rehab," where Tuttle comes out as Trixie, a trans woman. While presented with the show's characteristic humor, the narrative ultimately supports Trixie's journey of self-acceptance and living authentically, with Stan becoming an ally. The portrayal is affirming of trans identity.

The show frequently satirizes the hypocrisy, bigotry, and self-righteousness of characters who identify as Christian, particularly Stan Smith. While satirical, the narrative often links these negative traits to the adherents' (often superficial) faith, reinforcing a problematic portrayal.

American Dad! often critiques Islamophobia and xenophobia, particularly through Stan's initial prejudices which are frequently challenged and proven wrong by the narrative. The show positions the audience to sympathize with Muslim characters and condemn bigotry against them.