The royal couple will also have a private lunch with Italian president Giorgio Napolitano.
The one-day trip in April was originally planned for last year but was postponed when the Queen fell ill.
The BBC's royal correspondent Peter Hunt said the Queen's "relaxed" meeting with the Pope will be at his residence.
He added that the Queen would not wear black or a veil.
In a short statement, the palace said: "The Queen, accompanied by the Duke of Edinburgh, will visit Rome on Thursday 3 April.
"Her Majesty and His Royal Highness are visiting at the invitation of the president of Italy, President Napolitano.
"The Queen and the Duke of Edinburgh will visit the presidential palace where they will attend a private lunch hosted by the president.
"Following the private lunch Her Majesty and His Royal Highness will have an audience with His Holiness Pope Francis at the Vatican."
Source: BBC News
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