The Rookie features openly gay main character Aaron Thorsen, whose identity is depicted with dignity and normalcy. His relationships and personal struggles are handled respectfully, affirming his worth without relying on stereotypes. Other LGBTQ+ characters are also integrated incidentally, contributing to a net positive and validating portrayal.

The Rookie features a transgender character in Season 5, Episode 17, whose identity is handled with respect and empathy. The episode critiques misgendering and deadnaming, with a main character actively advocating for the victim's dignity and chosen identity. The portrayal is affirming, framing obstacles as external prejudice.

The show generally portrays Christianity as a source of personal comfort, moral guidance, and community for its characters. While individual characters may exhibit flaws, the narrative consistently treats the faith itself and its adherents with respect and sympathy, aligning with its virtues rather than critiquing them.