U.S. Highway 27 (US 27) in Florida is a north–south United States Numbered Highway. It runs 496.352 miles (798.801 km) from the Miami metropolitan area northwest to the Tallahassee metropolitan area. Throughout the state, US 27 has been designated the Claude Pepper Memorial Highway by the Florida Legislature. It was named after long-time Florida statesperson Claude Pepper, who served in both the U.S. Senate and House of Representatives. The stretch running from Miami to South Bay was originally designated the Thomas E. Will Memorial Highway by the Florida Legislature in 1937 when that portion was known as State Road 26 (SR 26). Thomas E. Will, the founder of Okeelanta, had worked for almost 20 years to get the state to build a road from Miami to the area south of Lake Okeechobee. For most of its length in the state, US 27 is a divided highway.

Claude Denson Pepper was an American politician of the Democratic Party. He represented Florida in the United States Senate from 1936 to 1951, and the Miami area in the United States House of Representatives from 1963 until 1989. He was considered a spokesman for left-liberalism and the elderly.