Rose joined a list of high-profile players that won't be at Muirfield this weekend after shooting 6-over 77 on Friday to finish on 10 over after two rounds.
The cut was made at 8 over, meaning Luke Donald (10 over), Rory McIlroy (12 over) and top-30 players Matteo Manassero (15 over) and Nick Watney (10 over) go home, too.
Rose is the first reigning U.S. Open champion to miss the cut at the British Open since Lucas Glover in 2009, with his decision to take three weeks off to digest his victory at Merion backfiring on a viciously difficult Muirfield course.
"I wasn't quite prepared to play the U.S. Open again," said the third-ranked Rose. "Golf humbles you all the time."
The bouncy fairways and lightning-fast greens at Muirfield reduced many of the world's best players to the role of a Sunday morning-hacker.
There was no let-up to the woes of McIlroy, with the two-time major winner and world No. 2 shooting 75 to go with his first-round 79. He greeted a birdie 4 at No. 17 with a fist-pump, entirely in gest.
"That was a very big putt for me," he said, laughing.
"It was a bit like (there was) sympathy for me out there."
Nicolas Colsaerts of Belgium six-putted on the bone-dry 15th green — and missed the cut by a stroke on 9 over.
Multiple major winners Tom Watson (11 over) and Vijay Singh (12 over) disappeared, as did Jim Furyk (10 over) and Rickie Fowler (12 over).
And there was no triumphant return to Muirfield for Nick Faldo, who won here in 1987 and 1992. He departed on 15 over.
Unlike Faldo, there will be plenty more opportunities for Rose at his home Open.
"It's just a bad week," Rose said. "To be honest, there are so many variables out on that golf course, I'm not even going to take it seriously, give myself a hard time. I'm not looking too deeply into it.
"The preparation has been unusual for me, having three weeks off. That was always going to be the slight unknown, I'm afraid ... I spent 2 weeks with my kids. Had a life. And enjoyed it. No regrets."