Gasperini entered the Juventus youth system at the age of 9; during his stay at the youth system, he won an Allievi Nazionali championship and was in the Primavera squad, which included Paolo Rossi and Sergio Brio, that placed runner-up in 1976 behind Lazio. After having played a handful of Coppa Italia matches with the first team, he was loaned to Reggiana and then sold to Serie B club Palermo in 1978. He stayed five seasons at Palermo, all in Serie B, but reached a Coppa Italia final in 1979, then lost to Juventus.
After two seasons with Cavese (Serie B) and Pistoiese (Serie C1), Gasperini moved to Pescara, where he finally gained his first opportunity to play in Serie A after the promotion in 1987. He made his Serie A debut in a home match against Pisa, ended in a 2–1 victory which featured a goal of his. In 1990, he left Pescara to join Salernitana, and retired in 1993 at the age of 35 after two seasons with Vis Pesaro.
In 1994 Gasperini returned to Juventus's youth system, this time as a coach. He was initially coach of the Giovanissimi (U-14) for two years, followed by two other years with the Allievi (U-17). In 1998, he became the manager of the Primavera (U-20) squad.
In 2003, he left Juventus to become head coach of Serie C1 club Crotone, where he readily guided his team to promotion in Serie B after play-offs. He stayed at Crotone for two more season in Serie B; he was sacked during the 2004–2005 season but appointed back soon later.
From 2006 he was head coach of ambitious club Genoa, and led his side to a promotion to Serie A in his first season with the rossoblu. In the 2008–2009 season, Gasperini led Genoa to fifth place of Serie A, the highest placement for the team in 19 years, thus securing a UEFA Europa League spot, relaunching players like Diego Milito and Thiago Motta in a 3–4–3 formation and a particularly spectacular football style that was praised throughout Italy, so much so that José Mourinho, manager of Serie A champions Internazionale, stated Gasperini was the coach who put him in greatest difficulty. However, a poor start in the 2010–11 season, with 11 points in 10 games despite popular signings such as Luca Toni, Rafinha, Miguel Veloso and Kakha Kaladze, caused Gasperini's dismissal from his coaching post on 8 November.
On 24 June 2011, Massimo Moratti confirmed that Gasperini would replace Leonardo as the manager of Internazionale. However, on 21 September 2011, Gasperini was sacked after a dismal run of five winless games, including four defeats.
Gasperini began his spell at Inter with a 2–1 loss against crosstown rivals Milan in the 2011 Supercoppa Italiana. In the first Serie A league game, Inter were then surprised by a caretaker-headed Palermo in a 4–3 defeat in Sicily, then followed by a scoreless home draw with Roma.
A 1–0 home defeat to Trabzonspor in the Champions League made matters worse, and Moratti sacked Gasperini after a shock 3–1 defeat to Serie A newcomers Novara.
On 16 September 2012, Gasperini was announced as the new manager of Palermo, a former team of his as a player, taking over from Giuseppe Sannino.
On 4 February 2013, he was dismissed from his post following a 2–1 loss at home to Atalanta.
On 24 February 2013, Gasperini was rehired as the Palermo manager, replacing Alberto Malesani after three games in charge. On 11 March 2013, Gasperini was again removed from the post, this time by Giuseppe Sannino.
On 29 September 2013, Genoa announced to have rehired Gasperini after almost three years since his previous spell ended.
On 14 June 2016, Gasperini was appointed manager of Atalanta.As of 20 September 2017