The 'Suit and Tie' star will be the headliner at DIRECTV's 'Super Bowl Saturday Night' benefit in New Orleans on February 2, which will raise money for the Shriners Hospital for Children.
Timberlake's charity concert will be co-hosted by mogul Mark Cuban and is to feature a DJ set from Questlove of The Roots.
He is expected to perform tracks from his upcoming album The 20/20 Experience, his first release since 2006.
Timberlake's new single 'Suit & Tie' - which also features Jay-Z - set US radio records by attaining the highest amount of first-week plays.
The 20/20 Experience was recently hailed as a "great" album by Justin Bieber, who also congratulated Timberlake on Twitter for his return to music.
The singer is expected to release The 20/20 Experience later this year.
> Justin Timberlake records new album 'The 20/20 Experience' - pictures
Listen to Justin Timberlake's 'Suit and Tie' below: