The novel follows the story of Genly Ai, a human native of Terra, who is sent to the planet of Gethen as an envoy of the Ekumen, a loose confederation of planets. Ai's mission is to persuade the nations of Gethen to join the Ekumen, but he is stymied by a lack of understanding of their culture. Individuals on Gethen are ambisexual, with no fixed sex; this has a strong influence on the culture of the planet, and creates a barrier of understanding for Ai.

The Left Hand of Darkness was among the first books in the genre now known as feminist science fiction and is the most famous examination of androgyny in science fiction.

A major theme of the novel is the effect of sex and gender on culture and society, explored in particular through the relationship between Ai and Estraven, a Gethenian politician who trusts and helps Ai. When the book was first published, the gender theme touched off a feminist debate over the depiction of the ambisexual Gethenians.

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