The Moldovan champions were narrowly edged out by last year's finalists Roma on matchday one, while Slavia won convincingly in Geneva against Servette.
Match preview
Slavia's 2-0 win in Switzerland two weeks ago solidified their status as favourites to progress along with Roma in this section.
The Slaviste have reached European quarter-finals in three of the last five seasons, but will hope to put things right after a poor showing last season.
Failing to progress from a relatively weak Europa Conference League group containing Sivasspor, CFR Cluj and Ballkani was hugely disappointing, but they have made the perfect start this time around with a win on the road.
However, they are winless in the Czech top flight since that win on the continent, after slipping up against Teplice at the weekend.
Jindrich Trpisovsky's side were relatively fortunate to escape with a 0-0 draw too, considering their hosts missed a first-half penalty.
Falling two points behind Sparta Prague, who had a tough test against Viktoria Plzen which they came through, will be disappointing, so Trpisovsky will hope his side can get back on track in Europe.
Their other league clash since the trip to Geneva saw them draw in the Prague derby with Sparta, but despite going winless in those two league games, Slavia have lost just three games in their last 47.
Slavia's home record in Europe is something to marvel at too, as the Czech side have lost just three of their 23 at the Fortuna Arena on the continent.
The home support may have revenge on their minds too coming into this clash, as Sheriff were the side who surprisingly knocked them out of Champions League qualifying back in 2009.
It would be a similar level of shock should Sheriff repeat the feat and claim something from this encounter.
Sheriff only progressed on away goals in that tie, as they are still yet to ever win a match on Czech soil - drawing three and losing twice.
Finishing in the top two would also be something of a turn up for the books, as Sheriff have never progressed directly through the groups in Europe.
Another defeat here after losing to Roma on matchday one may see Sheriff have to settle for a battle for third in the section.
That is what they managed last season, and saw them drop into the Europa Conference League where Sheriff went on to win their first-ever European knockout tie - beating Partizan Belgrade.
This competition has often provided a much tougher test though, as the Moldovan champions have won just two of their last 16 in the Europa League.
Slavia Prague Europa League form:
W
D
W
L
W
Slavia Prague form (all competitions):
L
W
W
W
D
D
Sheriff Tiraspol Europa League form:
W
D
D
W
L
Sheriff Tiraspol form (all competitions):
Advert - content below:
Team News
Slavia will have some injury problems in midfield, with three first-team squad members currently absent.
Petr Sevcik, Jakub Hromada and David Pech all remain out, so manager Trpisovsky must call upon some fringe players once more to fill the gaps.
Lukas Masopust has found a new lease of life remarkably as a right centre-back in recent weeks, despite naturally being a right-winger.
The 30-year-old scored on matchday one from that position, and will likely remain there despite the return of Jan Boril, who has dealt with injury and suspension recently.
Sheriff will be without Cape Verde international Joao Paulo after he was sent off late on against Roma on matchday one.
Gaby Kiki may also be absent after he missed the weekend win over Balti, with Rasheed Akanbi, Abou Ouattara and Vinicius Paiva all also ruled out.
Amine Talal started in the 4-0 win at the weekend, scoring twice, but with the game already sealed, he was withdrawn at half-time to keep him fresh for this one.
Fellow attacker Luvannor could come in from the side which lost to Roma too thanks to his excellent recent form which has seen him score five goals from six starts since mid-August.
Slavia Prague possible starting lineup:
Mandous; Masopust, Holes, Ogbu; Tomic, Jurasek, Dorley, Dumitrescu; Doudera, Tijani, Wallem
Sheriff Tiraspol possible starting lineup:
Koval; Tovar, Apostolakis, Garananga, Artunduaga; Zohouri, Ademo, Ankeye; Luvannor, Talal, Ngom Mbekeli
We say: Slavia Prague 2-0 Sheriff Tiraspol
Goals are not a common sight in Slavia games, even domestically where they often dominate and scored 98 goals in the league last season, as 10 of their last 12 outings have seen two goals or fewer.