When the Federal Communications Commission (FCC) lifted the "Freeze of '48"—the nationwide halt to reorganize TV frequencies—on April 14, 1952, the VHF channel 3 was assigned to the Kirksville, Missouri market.

[2] Hoping to defray the costs and risk of the new venture, in early 1953 the Kirksville group joined with another led by James J. Conroy, owner of KBIZ in Ottumwa, Iowa.

However, by time of sign-on the Kirksville group felt the venture held little hope for profitability and had sold all shares back to Conroy.

[2] For much of its early history KTVO was a primary CBS affiliate, although its single market status allowed it to cherry-pick the most popular programming from NBC and ABC.

On January 7, 1964, Conroy sold KTVO to the Post Corporation, a media conglomerate of newspapers and broadcasting properties based in Appleton, Wisconsin.

The new owners renovated and expanded the KTVO facilities in Ottumwa, shared for the station's first twenty years with then co-owned KBIZ-AM 1240.

[2] On June 1, 1968, KTVO switched primary affiliation to ABC, although the station continued carrying a few CBS and NBC shows until 1974.

This new height and location would allow KTVO's signal to reach the lucrative Quincy, Illinois–Hannibal, Missouri market while still serving its two original cities of license.

However, due to FCC duopoly regulations, KYOU was transferred to Ottumwa Media Holdings, LLC and entered into a local marketing agreement (LMA) with KTVO.

KTVO had been serving the Quincy–Hannibal–Keokuk, Iowa market as the default analog ABC affiliate on cable with some locations being able pick up its over-the-air signal.

On August 28, 2007, sister station KHQA (which had served as CBS' affiliate of record in the Kirksville market after 1974) announced that they would launch a new second digital subchannel to offer ABC programming beginning September 30.

On November 7, 2024, it was announced that KTVO and KHQA would merge their news departments into a new operation called Tri-State Trusted, which would cover both the Ottumwa–Kirksville and Quincy–Hannibal–Keokuk markets.

A list of fake names many considered crude or obscene (i.e. Dixie Normus and Craven Moorhead) was submitted to its weekday morning show birthdays/anniversaries segment.