Tuskegee ( tuh-SKEE-ghee) is a city in Macon County, Alabama, United States. General Thomas Simpson Woodward, a Creek War veteran under Andrew Jackson, laid out the city and founded it in 1833. It became the county seat in the same year and it was incorporated on February 13, 1843. It is the most populous city in Macon County. The population was 9,395 at the 2020 census, and was estimated to be 8,765 in 2023.

Tuskegee has been important in African-American history and highly influential in United States history since the 19th century. Before the American Civil War the area was developed for cotton plantations, dependent on enslaved African-American people.

After the war many freedmen continued to work on plantations in the rural area, which was devoted to agriculture, primarily cotton as a commodity crop. In 1881, the Tuskegee Normal School (now Tuskegee University, a historically black college) was founded by Lewis Adams, a former slave whose father, Jesse Adams, a white slave owner had allowed him to be educated. Its first founding principal was Booker T. Washington, who developed a national reputation and philanthropic network to support education of freedmen and their children.

In 1923, the Tuskegee Veterans Administration Medical Center was established, initially for the estimated 300,000 African-American veterans of World War I in the South, when public facilities were racially segregated. Twenty-seven buildings were constructed on the 464-acre campus.

The city was the subject of a civil rights case, Gomillion v. Lightfoot (1960), in which the United States Supreme Court ruled that the state legislature had violated the Fifteenth Amendment in 1957 by gerrymandering city boundaries as a 28-sided figure that excluded nearly all black voters and residents, and none of the white voters or residents. The city's boundaries were restored in 1961 after the ruling.