Sano went 1-for-4, with his first big-league hit being an infield single in the Twins' 2-0 victory over the Royals.
The 22-year-old third baseman from the Dominican Republic took the roster spot of first baseman/designated hitter Kennys Vargas, who was optioned to Double-A Chattanooga on Wednesday following the Twins' 2-1 loss to the Reds.
The Twins made Sano's call-up official Thursday and said he would wear No. 22. Sano celebrated his promotion on his Twitter account.
Sano served as the designated hitter with Trevor Plouffe already manning third base for the Twins and batted sixth.
Sano hit two home runs in 10 at-bats in spring training, with three walks in six games, but was sent to the minors before the season. He was bound to start in the minors, after missing last season while recovering from ligament-replacement surgery on his right elbow.
In 66 games at Chattanooga this season, Sano was hitting .274 with 15 home runs and 48 RBIs. In 2013, Sano combined to hit 35 homers in 123 games in Class A and Double-A.
Sano is the second top offensive prospect recalled by the Twins this season. Outfielder Byron Buxton was recalled last month but is now on the disabled list with a sprained left thumb. Buxton hit .189 in 11 games with five runs, a double, a triple and a steal plus two outfield assists before he went on the DL.
Sano was the No. 15 prospect in ESPN Insider Keith Law's Top 100 prospects for the 2015 season. Buxton was ranked No. 2.
The Twins have struggled to find consistent power in the middle of their lineup, particularly at designated hitter. They got off to a surprising start and were in first place in May. But they have gone 12-18 in their past 30 games, mostly because of poor offense.
The Twins trail the first-place Royals by 3½ games in the AL Central standings.
The Associated Press contributed to this report.