The Old Lady have appointed previous Italian national team manager the veteran manager to lead the team for the remainder of the campaign.
Spalletti replaces Igor Tudor, who was let go post the Lazio setback on Sunday.
Tudor had joined the club in March filling the shoes of Thiago Motta but left after seven months at the end of an eight-match winless run.
Spalletti led Napoli to the Serie A title in 2022-23 prior to assuming the Italy role, which missed out on the 2022 World Cup under former boss Roberto Mancini.
He secured their spot in Euro 2024 exiting in the last 16 stage against Switzerland.
However, Spalletti was sacked by Italy in June following a loss to Norway in World Cup qualifying, as Italy faced the possibility of not making a third World Cup in a row.
Spalletti has also managed Inter Milan, Roma and Udinese in Serie A and secured two Russian Premier League titles with Zenit St Petersburg.
Massimo Brambilla served as interim manager for Juventus during the 3-1 victory over Udinese on Wednesday moving them to seventh position in the league - six points behind leaders Napoli after nine games.
In the Champions League, they sit 25th in the league phase, having lost once and drawn twice in their opening three matches.
