The late will make a cameo appearance in the upcoming children's film, Paddington in Peru, to be released on November 8.
The brief appearance will be a photo of her having afternoon tea with the beloved animated bear, which has been approved by .
The head of global productions at StudioCanal, the film's distributor, said the decision to include the late monarch came "with the consent and agreement of the royal household".
It will be an emotional cameo for fans, in a short film for her 2022 celebrations, just months before her death.
on September 8, 2022, many tributes left outside her residences featured nods to the bear and her skit with him.
Rosie Alison, the film's co-producer, told Variety magazine: "[The royal family] were actually very happy for it to happen.
"But we don't like to make a big deal of it, because Paddington's obviously a very modest fellow."
In the original video for her Platinum Jubilee celebrations, in Buckingham Palace, during which the late Queen whipped a marmalade sandwich out of her bag, and said to Paddington: "I keep mine in here - for later."
The sketch touched the hearts of so many people that after her death a few months later more than 1,000 Paddington Bear soft toys were left outside royal residences. released a picture of Queen Camilla surrounded by some of the bears at Clarence House.
The Peruvian-born bear, who congratulated the late Queen on her remarkable reign during the sketch, will be seen heading back to his homeland, Peru, in the latest film. He gets into unexpected adventures in the Amazon rainforest and on mountain peaks, meeting interesting characters along the way.
Fans have rushed to social media to express their joy at seeing the late Queen's return to film. One said: "This is lovely."
Another wrote: "Lovely, thoughtful, human gesture."
New cast members include actors; as the Reverend Mother, and Antonio Banderas as Hunter Cabot.
The returning cast includes Ben Whishaw voicing Paddington, alongside Julie Walters, Jim Broadbent, Imelda Staunton, Hugh Bonneville and Emily Mortimer.
Paddington Bear was created by author Michael Bond in 1958 after he was inspired by the plight of refugee children arriving in the UK.
Paddington In Peru will be in cinemas on Friday, November 8.
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