William Blake, who died in 1827, described the painting Ancient of Days as "the best I have ever finished".
The artist and poet, whose talents were not fully appreciated while he was alive, had professed to wanting to see his work adorn the walls of churches and buildings.
Now to mark his birthday, Ancient of Days will illuminate the south side of the dome of St Paul's.
Martin Myrone, senior curator of pre-1800 British art at Tate Britain, said: "He had a lifelong ambition to see his paintings painted on a large scale....
"It was never going to happen. It was a pipe dream.
"Ancient Of Days is the very last thing he was working on. It's the thing he was colouring and painting in the very last days of his life.
"He is said to have stated it is the best thing he has ever done. It's an extraordinary work of art."
Ancient of Days was created as an illustration for Blake's book Europe a Prophecy and suggests his own interpretation of Christianity.
"It represents an idea of a perhaps rather oppressive, law giving judgemental God," Mr Myrone said.
"Blake was a very committed Christian but he had his own interpretation of religion....
"What you see is a realisation of a judging God looming from the heavens."
Born in Soho, London on 28 November 1757, William Blake was a writer, engraver and painter, whose prodigious talent went largely unrecognised during his lifetime.
He is now considered a seminal figure in the history of the poetry and visual arts of the Romantic Age.
Art critic and columnist Jonathan Jones has described Blake as "far and away the greatest artist Britain has ever produced". In 2002 he was placed at number 38 in the BBC's poll of the 100 Greatest Britons.
The projection of Ancient of Days onto St Paul's is part of a collaboration between the cathedral and Tate Britain, which is holding an exhibition of his works.
Dr Paula Gooder, Chancellor of St Paul's Cathedral, said: "This collaboration is made even more special because of the memorial in our crypt to William Blake.
"We hope that the projection of this iconic image will be an inspiration to all who see it."
The projections will run from 28 November to 1 December, from 4.30pm to 9pm each evening.