The Argentine, along with Weston McKennie, picked up an injury in the 3-1 victory against Sassuolo at the weekend and had to be replaced before half-time.
That will mean a recall for Alvaro Morata, who was only given four minutes of game time in the league, with the Spaniard coming in to partner Cristiano Ronaldo in attack.
The former Real Madrid man is returning from injury and isn't fully fit but could be asked to start due to the absence of Dybala.
McKennie's injury was even quicker against Sassuolo as he lasted just 19 minutes before having to be replaced. Expect Dejan Kulusevski to replace the United States international on the left-hand side of midfield.
Unfortunately for the Bianconeri, they will also be without Matthijs de Ligt, Alex Sandro and Juan Cuadrado as the trio as still in isolation after previously testing positive for coronavirus.
Gianluigi Buffon will likely come in to replace Wojciech Szczesny in goal.
Even though Pirlo has plenty of players missing, he still should be able to deploy his preferred 4-4-2 formation.
Danilo can feature at right-back, with Leonardo Bonucci and Merih Demiral and youngster Gianluca Frabotta at left-back.
Federico Chiesa also picked up a knock against Sassuolo and might not be chanced in the cup, with Juve likely to have one eye on the eye match against Inter this weekend.
That could mean a start on the right for Federico Bernardeschi.
In the middle, Adrien Rabiot could come in for Rodrigo Bentancur, though Aaron Ramsey is pushing for a start too after coming off the bench to score in Serie A.