Moore suffered the injury in the final minute of Saturday's Elite Eight win over Houston. He is out indefinitely.
"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," coach Jay Wright said in a statement. "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.
"Justin's work ethic and dedication will help bring him through this recovery and we look forward to the day where we see him back on the court fully healthy."
Villanova faces 1-seed Kansas in Saturday's Final Four matchup.
Moore slipped while driving to the lane with 35 seconds left in Saturday's 50-44 win and had to be helped off the court. He did not take part in the postgame celebrations, with his teammates coming to see him at the end of the bench after the final buzzer sounded.
Moore went to the locker room on crutches during the trophy ceremony.
"I've been in that position before," Villanova star guard Collin Gillespie said after Saturday's game. "I know what it feels like. We don't know what the injury is yet. He still has to get checked. I don't ever want to see him by himself. He's done so much for his team, always guards the best player, is one of our best offensive players and will do anything for any one of our guys on either end of the floor.
"A lot of our guys ran right over to him because we're not in the position that we are without him. So we just wanted to go over to him and just appreciate what he does for us and know that we got his back."
Wildcats senior forward Jermaine Samuels said he recollected the injury to Gillespie, who tore a ligament in his knee last March.
"I still don't know what's necessarily going on, but I knew something wasn't right," Samuels said of Moore. "It kind of gave me a flashback to when the same thing happened to Collin last year, where it was like a numb type of feeling."
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 and earning second-team All-Big East honors. He was also a member of the All-South Region Team announced on Saturday after scoring 21 points in the first-round win over Delaware and 15 points in the Sweet 16 victory over Michigan.
With Moore out for the Final Four, senior guard Caleb Daniels will step into the starting lineup. The Wildcats' sixth man, Daniels had 12 points in 35 minutes against Houston. He averages 10.2 points and 3.8 rebounds this season, shooting 37.7% from 3-point range.
The biggest impact will be felt on Villanova's bench. Reserves Chris Arcidiacono and Bryan Antoine will see an increase in minutes. Arcidiacono has played six minutes in the past three games, while Antoine didn't play against Houston but saw a combined eight minutes of action against Michigan and Ohio State.