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The Duke and Duchess of Sussex will be questioned as part of a US defamation case brought by Meghan's half-sister.
This comes after a motion to stop depositions in the civil suit from taking place was dismissed by a judge.
Samantha Markle is suing Meghan for "defamation and injurious falsehoods" following the royal couple's interview with Oprah Winfrey in 2021.
She alleges she was defamed when Meghan "falsely and maliciously" said she was "an only child".
The much-anticipated interview with Winfrey saw the couple discuss their families, royal life and mental health.
The duchess' half-sister claimed, in a filing submitted in March, that Meghan made "demonstrably false and malicious statements" to a worldwide audience.
This case comes after the publication of Prince Harry's memoir Spare in January and the couple's Netflix series last year.
A deposition is a formal statement of evidence required to be taken of a witness or party to litigation by a court.
The duchess' motion to stop depositions in the case from taking place was dismissed on Tuesday by Florida judge Charlene Edwards Honeywell.
The documents, obtained by PA News agency, stated: "Defendant Markle does not show that unusual circumstances justify the requested stay, or that prejudice or an undue burden will result if the court does not impose a stay.
"Defendant Markle does not satisfy the high standard required to stay discovery pending the resolution of a dispositive motion."
Samantha Markle's original complaint, also obtained by PA, stated the comments made by the duke and duchess during their interview with talk show host Winfrey had reached "roughly 50 million people in 17 countries".
She alleged, in the filing, that she was defamed by Meghan when the duchess "falsely and maliciously" said she was "an only child".
"Plaintiff - who suffers from multiple sclerosis and is confined to a wheelchair - brings this action for defamation based on demonstrably false and malicious statements made by her half-sister to a worldwide audience, including roughly 50 million people in 17 countries who watched the Oprah Winfrey interview with the Defendant, Meghan Markle, and her husband, Prince Harry of England," the filing stated.
"Meghan - who was featured with Prince Harry on the cover of Time Magazine's annual feature on 'The World's Most Influential People' published and disseminated false and malicious lies designed to destroy Plaintiff's reputation and which have subjected Plaintiff to humiliation, shame and hatred on a worldwide scale."
It was also added in the filing that Meghan had used "the powerful resources of the royal family's public relations operation" to disseminate and spread "lies worldwide" about her half-sister and their father Thomas Markle.
"Defendant orchestrated the campaign to defame and destroy her sister's and her father's reputation and credibility in order to preserve and promote the false 'rags-to-royalty' narrative," the filing stated.
"[It was] a premeditated campaign to destroy their reputation and credibility so they could not interfere with or contradict the false narrative and fairy tale life story concocted by the defendant."