The temporary closure of the highway between the Watterson Expressway and downtown is set for June and July 2026.
The crash was reported in the southbound lanes, near the Louisville Muhammed Ali International Airport.
The woman was attempting to cross the street when she was hit.
INDOT said the highway is expected to be closed for several hours.
Work is taking place until 3 p.m. on Thursday, July 27, and from 9 a.m. to 3 p.m. on Friday.
A busy section of Interstate 65 in Louisville is pitch black, with no working streetlights for more than two miles.
Summer is the season for road work, and one of the latest projects to be announced is on Interstate 65 near the border of Bullitt and Hardin counties.
The Spring Street ramp to I-64 Westbound will be closed on Friday, June 2 from 8 a.m. to noon, weather permitting.
The crash is causing significant delays in the westbound lanes of I-264.
The road is closed coming from Jeffersonville into Clarksville on the overpass of I-65, according to Clarksville Police dispatch.