According to Harmonix's lead designer, Chris Foster, the idea of The Beatles: Rock Band came about during a chance encounter between MTV president Van Toffler and Dhani Harrison, son of George Harrison. Dhani, having been familiar with Rock Band, suggested a game based on The Beatles; both Dhani and Toffler considered the concept an unlikely possibility. Nonetheless, the meeting spurred Dhani into further discussions with Harmonix's president, Alex Rigopulos, sharing his ideas for the game. At the same time, Dhani helped to introduce the Rock Band concept to Apple Corps and its key shareholders Sir Paul McCartney, Ringo Starr and Yoko Ono Lennon.
Harmonix subsequently created a simple demonstration of the game that included examples of music and artwork. With the creation of the demo, the remaining members of The Beatles as well as Ono Lennon and Olivia Harrison were brought in as creative partners for the game. They helped decide the songs and venues that would appear in the game, as well as provided feedback on the artwork, character representations, and storyboards for animation sequences. The development team was able to bring Giles Martin aboard as the game's music producer; Martin having previously served as co-producer for The Beatles Love project. Giles was able to assure multitrack capabilities to The Beatles' master recordings, which is essential in allowing individual audio feedback to each player — for example, missing a note on the guitar track of a song will temporarily quiet the guitar audio. Harmonix performed very little additional remixing upon receipt of these remasters; in some cases, three different guitar parts—lead, solo, and rhythm—were brought into a single cohesive guitar part, slightly raising the volume of the specific guitar track that was used in note tracking to make it easier for the player to follow in the game. The ability for up to three players to sing vocal harmonies, a feature not present in previous Rock Band games, was designed and implemented as an optional feature so as not to be overwhelming to players.
The game will feature a visual and musical history of The Beatles and will include the United Kingdom-released versions of songs from their albums Please Please Me through to Let It Be. The game will "feature different periods of the band — you get early days, Liverpool, then psychedelic and on from there," according to McCartney. Dhani Harrison has stated the game will include "stuff that has never been heard, never been released." Gamestop's vice-president of sales, Bob McKenzie, noted after seeing a demonstration of the game, that the game includes "some of the cutscenes and some of the added material from the band and from that era." Some of the new material includes band chatter and instrument tunes taken from recorded performances, and are played during the loading screens and intros and outgos of songs

The challenging levels of amusement and gamesmanship:
"And Your Bird Can Sing" Budokan
"Back in the U.S.S.R." Abbey Road Studios / Dreamscape
"Birthday" Abbey Road Studios / Dreamscape
"Can't Buy Me Love" Ed Sullivan Theater
"Day Tripper" Budokan
"Dig a Pony" Rooftop Concert
"Do You Want to Know a Secret" Cavern Club
"Eight Days a Week" Shea Stadium
"Get Back" Rooftop Concert
"Here Comes the Sun" Abbey Road Studios / Dreamscape
"I Am the Walrus" Abbey Road Studios / Dreamscape
"I Feel Fine" Shea Stadium
"I Saw Her Standing There" Cavern Club
"I Want to Hold Your Hand" Ed Sullivan Theater
"I Wanna Be Your Man" Ed Sullivan Theater
"I've Got a Feeling" Rooftop Concert
"Octopus's Garden" Abbey Road Studios / Dreamscape
"Paperback Writer" Budokan
"Revolution" Abbey Road Studios / Dreamscape
"Sgt. Pepper's Lonely Hearts Club Band" / "With a Little Help from My Friends"a Abbey Road Studios / Dreamscape
"Taxman" Budokan
"Twist and Shout" Cavern Club
"Within You Without You / Tomorrow Never Knows"b Abbey Road Studios / Dreamscape
"Yellow Submarine" Abbey Road Studios / Dreamscape
Four new instrument peripherals modeled after those used by The Beatles members will be introduced with the game. These include a Rickenbacker 325 guitar, a Gretsch Duo Jet guitar, a Höfner bass and a Ludwig drum set.
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