Starting from I-65 in the southern part of Louisville, it runs through Jefferson County, Kentucky, crosses the Ohio River on the Lewis and Clark Bridge into Indiana, meets I-65 for a second time, and then proceeds westbound to terminate at the I-64 interchange.

[citation needed] Likewise on the Indiana side, the stretch from I-65 to the bridge, while designated as I-265 is signed as State Route 265 (SR 265).

[citation needed] I-265 in the US state of Indiana presently runs 13.11 miles (21.10 km) from I-64 at the western edge of New Albany to the Lewis and Clark Bridge near Utica.

The Gene Snyder Freeway, in which KY 841 and I-265 overlap for 24.48 miles (39.40 km) between I-65 and the Indiana state line, has seen an increase in serious accidents.

The freeway was legally rechristened in June 2019 to I-265 after a new 7.7-mile (12.4 km) section of highway that includes the Lewis and Clark Bridge (known as the East End Bridge during planning and construction) fully opened to traffic on December 18, 2016, however the SR 265 signs along the road and the exits still remain and have not been updated to I-265 signage, while the Kentucky portion of the new roadway likewise has not had I-265 signage added as of yet.

In early 2023, Indiana DOT announced plans to redesignate as well as renumber the exits of the freeway beginning in the fall of 2023 and with completion expected over the next two years.

In late 2005, members of the Louisville Metro Council proposed a committee to begin planning a western bridge over the Ohio River to link the southwestern end of the highway in Kentucky to Indiana.