In a preview clip of Apple Music's "Carpool Karaoke: The Series" above, MacFarlane takes on the role of Seymour while Grande plays Audrey.
The episode will air Tuesday night on Apple Music as part of a 20-episode spinoff series of James Corden's creation.
Unlike the "Carpool Karaoke" segments featured on Corden's "The Late Late Show," he will not host the series regularly. He will, however, make a few cameos. The Apple series is actually hostless so the focus is 100 percent on the singing celebrities.
Grande got her start performing in Jason Robert Brown's "13" on Broadway. She also played Penny Pingleton in the NBC live television "Hairspray Live!" Although MacFarlane is most known for his presence on television, he often incorporates musical theater into his characters.