Organisers have made the decision to stage a special event after having to cancel the festival for a second year due to the ongoing Covid-19 pandemic.
Performers will take to the stage at various landmarks around Worthy Farm in Somerset, including the Pyramid field and the stone circle, during the five-hour event.
Damon Albarn, Kano, Michael Kiwanuka, and Idles are also on the line-up, while organiser Emily Eavis also teased some "very special guest appearances and collaborations" will be taking place.
Paul Dugdale, who shot Ariana Grande and Taylor Swift's world tours for Netflix, is in charge of directing the livestream, which will see the musical performances linked by "a spoken word narrative, written and delivered by some very special guests."
Tickets are on sale now, with proceeds going towards Oxfam, Greenpeace and WaterAid, as well as helping to "secure the Festival's return in 2022."
In addition, a commemorative poster of the livestream gig is to be sold in support of the stage crew workers who have been struggling to make ends meet during the global health crisis.
For tickets, visit: glastonburylivestream.seetickets.com.
The four-day bash in Tennessee was initially set for June, 2021, but was recently moved to 2 to 5 September due to ongoing COVID concerns but, when the event returns, fans will be treated to an all-star line-up, which also features Lana Del Rey, Janelle Monae, Tame Impala, and Tyler, the Creator.
Other acts on the bill for the 20th anniversary include Run the Jewels, Young Thug, Deftones, My Morning Jacket, G-Eazy, Phoebe Bridgers, Jason Isbell & The 400 Unit, Lil Baby, Deadmau5, and Leon Bridges.
In a statement celebrating the line-up news, Governor Bill Lee says, "It's exciting to see Tennessee stages come back to life in time to celebrate the 20th anniversary of this internationally acclaimed festival."
"Fans are ready to gather together and celebrate their shared love of music once again. We welcome them back for a full Bonnaroo and what is sure to be a truly unforgettable event!"
Tickets for the festival, which had its in-person gigs cancelled last year (20) due to the coronavirus pandemic, are on sale now.