Glasgow Warriors will begin the new competition on the road, as Danny Wilson's men travel to Benetton on the opening weekend.
The Warriors then welcome Edinburgh to Scotstoun on the weekend of 7/8/9 May for the first of back-to-back fixtures with their cross-country rivals, before travelling to BT Murrayfield for the reverse fixture on the weekend of 14/15/16 May.
How the Guinness PRO14 Rainbow Cup will work
Changes to scheduling and format of the Guinness PRO14 Rainbow Cup have been made to limit travel where possible with the opening three rounds accounting for derby fixtures across Ireland, Italy, Scotland, South Africa and Wales.
Rounds four to six will see the four South African 'Super' franchises play their away games in Europe and those fixtures will be confirmed once all appropriate travel approvals have been confirmed by the relevant authorities and governments.
This format means that no teams from Europe will travel to South Africa for Guinness Rainbow Cup games.
A total of six rounds (eight games per round) will take place with all teams ranked in a single league table. The top two teams will then play each other in the Guinness PRO14 Rainbow Cup Final on June 19.
PRO14 Rugby is currently consulting with its broadcast partners to select kick-off times and will confirm the scheduling as soon as possible.
David Jordan, Tournament Director, PRO14 Rugby said: "Everyone is glad that we can now put some certainty into the rugby calendar in what has been an unpredictable period. Planning for a cross-hemisphere competition where the situation has been so fluid has provided many challenges, but we believe we have found a solution that is practical and safe to deliver a unique competition that supporters can get excited about.
"Throughout the Guinness PRO14 season have liaised with our unions and their governments to ensure we are meeting the conditions set out for elite sport to operate. We are very grateful for that support, without which a cross-border competition such as ours could not have taken place."
2021 Guinness PRO14 Rainbow Cup fixtures
Round One: Benetton v Glasgow Warriors, Stadio Comunale di Monigo, w/o 23/24/25 April
Round Two: Glasgow Warriors v Edinburgh, Scotstoun Stadium, w/o 7/8/9 May
Round Three: Edinburgh v Glasgow Warriors, BT Murrayfield, w/o 14/15/16 May
Round Four: TBC, w/o 28/29/30 May
Round Five: TBC, w/o 4/5/6 June
Round Six: TBC, w/o 11/12/13 June
Final: TBC, w/o 18/19/20 June