Two-spirit (also known as two spirit or occasionally twospirited, or abbreviated as 2S or 2E, especially in Canada) is a contemporary pan-Indian umbrella term used by some Indigenous North Americans to describe Native people who fulfill a traditional third-gender (or other gender-variant) social role in their communities.

Coined in 1990 as a primarily ceremonial term promoting community recognition, in recent years more individuals have taken to self-identifying as two-spirit. Two-spirit, as a term and concept, is neither used nor accepted universally in Native American cultures. Indigenous cultures that have traditional roles for gender-nonconforming people have names in their own Indigenous languages for these people and the roles they fill in their communities.

The initial intent in coining the term was to differentiate Indigenous concepts of gender and sexuality from those of non-Native lesbians and gays and to replace the pejorative anthropological terms that were still in wide use. Although the term "two-spirit" has been controversial since its adoption, it has experienced more academic and social acceptance than the term berdache, which it was coined to replace. The government of Canada officially uses 2SLGBTQI+ as an alternative to the established acronym of LGBTQI+, sometimes shortened to 2SLGBT or a similar variant.

Early adopters stated that a two-spirit identity does not make sense outside of a Native American or First Nations cultural framework and its use by non-Natives is seen as a form of cultural appropriation.

The gender-nonconforming or third-gender ceremonial roles traditionally embodied by some Native American and Indigenous peoples in Canada that may be encompassed by modern two-spirit people vary widely, even among the Native individuals or cultures that use the term. Not all of these cultures have historically had roles for gender-variant people, and among those that do, no one Indigenous culture's gender or sexuality categories apply to all Native people.

The neologism two-spirit was developed over a series of five conferences, concluding in 1990 at the Third Annual Inter-tribal Native American, First Nations, Gay and Lesbian American Conference, held in Winnipeg. Credit for developing the term is usually given to several participants in the gatherings. The term was first developed and proposed in English, and later the Ojibwe translation niizh manidoowag was constructed and proposed to honor the language of the Indigenous peoples in whose territory the conference was being held.

Both the English and Ojibwe terms were coined at the 1990 conference, and are not found in the historical record. Two-spirit, in English or translated into any other language, is a general term for wider audiences, and is not intended to replace the traditional terms or concepts already in use in Indigenous cultures.

For early adopters, the term two spirit was a deliberate act to differentiate and distance themselves from non-Native gays and lesbians, as well as from non-Native terminology such as gay, lesbian, and transgender. Particularly offensive was the term berdache, which had previously been the preferred term among non-Native anthropologists to refer to Indigenous people who did not conform to standard European-American gender roles. Berdache, which means "passive partner in sodomy, boy prostitute", has always been offensive to Indigenous peoples. Journalist Mary Annette Pember (Red Cliff Ojibwe) and others have written that conference participants were motivated by the desire to coin a new term that could take the place of the outdated and offensive anthropological term.