Moore is set to undergo surgery this week and will be sidelined indefinitely.
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"This is a tough blow for all of us, not just because of the great player Justin is, but because of what he means to us as teammates and coaches," said Villanova head coach Jay Wright. "As one of our captains, Justin has been an integral part of our success. He helps our team in so many ways. Every game, he guards the opponent's best scorer and is one of the toughest rebounding guards we have had in our time at Villanova."
Moore slipped while driving to the lane with 35 seconds left in Saturday's game and had to be helped off the court.
Moore, a 6-foot-4 junior, had eight points and two rebounds in 37 minutes against Houston. He ranks second on the team in points and assists and third in rebounds, averaging 14.8 points, 4.8 rebounds and 2.3 assists.
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Senior guard Caleb Daniels is expected to step into the spotlight for Moore. The Wildcats' sixth man, Daniels had 12 points in 35 minutes against Houston. He averages 10.2 points and 3.8 rebounds per game this season, shooting 37.7% from 3-point range.
The Wildcats are going to their third Final Four in the past six NCAA Tournaments.
No. 2 Villanova will take on No. 1 Kansas on Saturday at 6:09 p.m.
--ESPN contributed to this report.