U.S. Route 53, or U.S. Highway 53 (U.S. 53), is a north–south U.S. highway that runs for 404 miles (650 km) from La Crosse, Wisconsin to International Falls, Minnesota. It is the primary north–south route in northwestern Wisconsin, serving as a vital link between I-94 at Eau Claire, Wisconsin and the Twin Ports of Superior, Wisconsin, and Duluth, Minnesota. The entire route from Eau Claire to the city limits of Superior is a four lane divided highway. The highway's northern terminus is at the Fort Frances–International Falls International Bridge in International Falls, Minnesota, at the Canada–US border. Its southern terminus is in La Crosse, Wisconsin, at U.S. Highway 14.

La Crosse is a city in and the county seat of La Crosse County, Wisconsin, United States. Positioned alongside the Mississippi River, La Crosse is the largest city on Wisconsin's western border. La Crosse's population was 52,680 as of the 2020 census. The city forms the core of the La Crosse–Onalaska metropolitan area, which includes all of La Crosse County and Houston County, Minnesota, with a population of 139,627.