Harry appeared in a comedy sketch that aired on Wednesday night, playing himself as he auditions to become a Hallmark movie "Christmas prince".
In the segment, the duke wandered through the studio, joking that he has become lost while searching for an audition for the "Gingerbread Christmas Prince saves Christmas in Nebraska".
Colbert asks why a real prince would want to star in such films, prompting Harry to reply: "You Americans are obsessed with Christmas movies, and you're clearly obsessed with royalty, so why not?"
When Colbert pushed back on the idea of an American "obsession," Harry quipped: "Really? I hear you elected a king," in an apparent reference to the "No Kings" protests directed at the Trump administration.
Referencing CBS's recent settlement with Trump, the duke joked that he would "do anything" to get the part, adding: "I'll record a self-tape, I'll fly myself to an audition, settle a baseless lawsuit with the White House - all the things you people in TV do."
Colbert replied: "Hey, I didn't do any of those things," to which Harry shot back: "Maybe that's why you're cancelled."
The sketch then transformed into a festive spectacle, with trees appearing and sleigh bells ringing as Harry urged Colbert to "just believe".
Colbert later told Harry he "got the role" of "official Late Show prince of Christmas" to cheers from the crowd.
The pair first teased the guest spot in a video featuring a viral Great British Bake Off sound bite on Wednesday.
Colbert's other guests were Academy Award nominee Michael Shannon and Irish actress and singer Jessie Buckley.
Wednesday's visit was the duke's second Late Show appearance, having last appeared in January 2023 to promote his memoir Spare.
Harry's first appearance on the show set a two-year viewership record and made international headlines.
The announcement of Wednesday night's appearance was also shared by the Duchess of Sussex, who previewed the clip on her Instagram story.
Harry and Meghan have been raising their children in Montecito, California, since announcing they were stepping back as working members of the royal family in January 2020.
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