The trail extends from the headwaters at Lake Itasca in Minnesota to near the mouth of the river in Venice, Louisiana.

The Mississippi River Trail route marker is based on a 2012 design that the Federal Highway Administration (FHWA) has allowed several states, including Illinois, Kentucky, and Tennessee, to use as U.S.

[2] However, the Mississippi River Trail's marker bears the abbreviation "MRT" in place of a numeric route designation.

[6] By the time the Minnesota Department of Transportation (MnDOT) successfully proposed designating the MRT as USBR 45, planning for the trail was already well underway.

After it leaves the park, the MRT winds north along the river, traversing county roads to the city of Bemidji.

From Bemidji, the MRT heads southeast along the Paul Bunyan Trail, a state-developed rail-trail conversion.

From Brainerd, the MRT follows county roads, some with marked bike lanes and some with paved shoulders, through the cities of Little Falls and St.

The Wisconsin section of the MRT begins at Prescott and continues along the eastern side of the river all the way to Illinois.

It continues south on SR 181 for several miles until it crosses over the Forked Deer River and into Lauderdale County, Tennessee.

The trail turns right onto Cooper Creek Road and then right onto U.S. Route 51 (US 51), crossing into Tipton County over the Hatchie River.

The trail travels along the ridge of the 3rd Chickasaw Bluff and passes through Meeman-Shelby Forest State Park.

Then the trail turns left onto South Circle Road (turning into East Circle Road), right onto Northaven Drive, and right onto SR 388 (North Watkins Street), passing through the Northaven area and crossing over the Loosahatchie River into the Frayser community of Memphis.

[11] Construction has been completed on a multi-use trail for pedestrians and cyclists along the north side of the Harahan bridge.