The 42-year-old Figueres native has served as the former Spain international's right-hand man since they assumed control of the Blaugrana's B team in 2007
Guardiola confirmed on April 27 that he will be stepping down as coach at the end of the season and the Blaugrana wasted little time in announcing his right-hand man of the past five years as his replacement.
The former Spain international's last official outing will come against Athletic Bilbao in the Copa del Rey final on May 25 but it has now been revealed that the 41-year-old will go head to head with Vilanova two days later.
Guardiola will take charge of one half of the Barca squad, and Vilanova the other, for a game TV3 and Radio Catalunya have organised to raise funds for a marathon charity event aimed at ending poverty.
"All of the football teams in our city are important but Barca are very important," Xavier Trias, the mayor of Barcelona, is quoted as saying by Formulatv.com.
"They will generate a huge media presence, and the fact that the first team have decided to step forward to play one another, along with the two coaches, is very significant. Thanks to the club and the two coaches."
One side will wear the Blaugrana's traditional colours for the match, which will be held at Palau Sant Jordi, while their adversaries for the evening will don their black away jersey.