Antonio Conte's men are favourites to progress to the tournament decider in Turin but they are going up against a side fresh from clinching a 33rd Primeira Liga title
The Italian giants last won continental silverware in the 1999 Intertoto Cup and know this year's final will be held at their home stadium in Turin.
That gives Juve extra motivation ahead of this two-legged clash and Antonio Conte's men will hope Benfica are still affected by their title celebrations.
Jorge Jesus' side were crowned champions of Portugal for the 33rd time thanks to a 2-0 win against Olhanense on Sunday.
Juventus are also set to win their own domestic title, which would give them a third successive Serie A success, but midfielder Claudio Marchisio is not worried about fatigue with so much at stake.
"Many players in this team have never won a European competition and Juventus haven't won a trophy in Europe for so many years," Marchisio told Uefa's official website.
"When you reach this stage of the competition motivation can make the difference.
"So even if we're coming to the end of the season and our legs are beginning to feel heavy, the trophy can give you that energy to give that bit extra."
Juventus are one of just two sides remaining in the Europa League who are undefeated – the other is Benfica – but both have only been involved since dropping out of the Champions League at the group stages.
This will be Juve's first semi-final appearance in Europe since 2003, when they eventually lost the Champions League final to Milan on penalties.
They have played Benfica over two legs in Uefa competition twice previously, with the Portuguese side winning both matches in the 1968 European Cup semi-final.
Juve exacted revenge in the 1992-93 Uefa Cup quarter-finals, with current coach Conte playing a part in the win that helped them on their way to winning the trophy.
Benfica's recent pedigree in the Europa League is particularly impressive, with the Portuguese club losing last season's final to Chelsea.
They also reached the semi-finals in the 2010-11 season, were involved in the quarter-finals the year before, and are trying to win an ambitious quadruple.
With the league title already sealed - and a final-four showdown against Porto in the Taca da Liga and a Taca de Portugal final with Rio Ave on the horizon – Benfica's hopes are still on track.
They are unbeaten in their last 12 home European encounters, while their record at Estadio da Luz in the Europa League is outstanding, with 15 wins and two draws coming from their 17 matches.
Benfica will be without winger Eduardo Salvio, who broke his arm in Sunday's win, while defender Silvio broke his leg in his side's quarter-final win against AZ Alkmaar.
Striker Carlos Tevez continues to battle a thigh niggle for Juve, but Andrea Barzagli (calf) did return in Saturday's 1-0 win over Bologna.
Simone Pepe was also part of the squad after recovering from a thigh injury, while Arturo Vidal shrugged off a knee complaint to feature off the bench.