Teams from 17 Premier League clubs and six Championship teams will be divided into three groups. From January onwards, they will be divided into another three groups based on their performance, which will lead to a knockout stage and final.
The national competition will launch on August 17 with Manchester City visiting Chelsea, and West Ham hosting Reading.
Each team will be allowed to field three outfield players over the age of 21 as well as an overage goalkeeper, in a similar style to Olympic football but with a lower age barrier.
"Each stage of a player's development is tailored to ensuring they have the best possible chance of progressing," Premier League chief executive Richard Scudamore said in a statement.
"The new Barclays Under-21 Premier League addresses one of the most critical points - the transition from academy football to the highest standards in the first team.
"We believe this competitive environment, where fans can also come along to watch the next generation of Premier League and international players, will help deliver more talented home grown boys to that next level."
The three groups are as follows:
- National Group 1 - Arsenal, Blackburn, Bolton, Everton, Norwich, Reading, West Brom, West Ham.
- National Group 2 - Aston Villa, Manchester United, Newcastle, Southampton, Stoke, Sunderland, Tottenham, Watford.
- National Group 3 - Chelsea, Crystal Palace, Fulham, Liverpool, Manchester City, Middlesbrough, Wolves.