Suleiman Abu Ghaith is accused of conspiracy to kill Americans and supporting terrorists in the immediate aftermath of the 9/11 attacks that killed 3,000 people.
The 48-year-old suspect from Kuwait faces life behind bars if convicted and is one of the most senior alleged Al-Qaeda members to face a US trial.
Abu Ghaith wore an oversized beige blazer and open-necked white shirt as he sat next to his defence lawyers and listened to a simultaneous translation.
Abu Ghaith is best known for appearing with bin Laden and the current leader of Al-Qaeda, Ayman al-Zawahiri, in al-Qaeda propaganda videos in September 2001.
Married to bin Laden's daughter Fatima, US prosecutors say Abu Ghaith worked for Al-Qaeda until 2002, when he fled Afghanistan after the US invasion for Iran.
The prosecution claims he was complicit in the December 2001 shoe bombing plot to bring down an airliner flying from Paris to Miami.
The defendant pleads not guilty to all three counts.
Highlights of the trial are likely to be two witnesses testifying by video link from Britain and Yemen.
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