Gonzalo Higuain had put Juventus in front in Saturday’s Serie A finale but Perotti set up Nikola Kalinic’s equaliser and put Roma ahead from the penalty spot before half-time.
The winger gave fifth-placed Roma a cushion early in the second half as Juventus succumbed to a first home Serie A defeat since a 1-0 reverse to Napoli in April 2018 – a run of 39 games.
It means champions Juve, who rested several players including Cristiano Ronaldo, finish the campaign with back-to-back losses before aiming to overturn a last-16 deficit against Ligue 1 Lyon.
Higuain turned in from close range just six minutes in after pouncing on Adrien Rabiot’s flick from Federico Bernardeschi’s corner.
Juve were undone by a corner themselves, though, as Kalinic nonchalantly converted Perotti’s delivery.
Gianluca Frabotta – making a first Juve appearance – dragged wide, while fellow youngster Riccardo Calafiori rifled home at the other end, only for the ball to have gone out of play from the preceding corner.
Calafiori then drew the foul from Danilo for Perotti’s penalty to find the bottom-right corner and put Roma ahead.
Perotti stretched the lead seven minutes into the second half, sweeping home inside the area – via a deflection off Merih Demiral – after collecting Nicolo Zaniolo’s perfectly weighted pass.
Aaron Ramsey had a with a 20-yard strike hit to the post with a sweet effort and Higuain nearly found the bottom-left corner .
But Roma had other chances themselves, with Davide Zappacosta shooting narrowly wide and Wojciech Szczesny made a couple of decent saves as Juventus faltered before a huge Champions League showdown.
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