Owen Farrell has been left out of England head coach Steve Borthwick's matchday 23 to face Ireland at the Aviva Stadium in Dublin on Saturday.
Farrell was due to feature in the Rugby World Cup warm-up fixture but has now been removed by Borthwick due to World Rugby's appeal against the disciplinary decision that saw his red card from England's match against Wales overturned, putting his World Cup hopes in renewed doubt.
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Borthwick said: "In my original planning for the Summer Nations Series and as part of our Rugby World Cup preparation, Owen Farrell was due to be selected to play in this Saturday's Test match in Dublin.
"However, in light of the fact that so much of Owen's training and match preparation time this week has been significantly affected and interrupted by the disciplinary process, I have not selected him in the matchday 23. Owen will travel to Ireland with our full support.
"Owen understands the situation, but is of course disappointed, as I am, that he is missing such a special game that he would otherwise have played in."
Farrell's hearing has now been confirmed for Tuesday, Six Nations Rugby saying: "Six Nations Rugby can confirm it has received an appeal from World Rugby in respect of Owen Farrell's independent disciplinary hearing decision.
"An independent appeal committee will hear the case early next week at a time and date to be confirmed.
"The chair and members of the committee will be confirmed in the coming days."
Speaking ahead of the match Andy Farrell, Ireland head coach and Owen's father, also came out strongly about the reaction to the overturning of his son's red card, calling it "absolutely disgusting".
"Whatever I say is probably flawed. When you're asking someone about their son, it's going to be flawed," said Farrell Sr.
"What I would say is that the circus around this is absolutely disgusting.
"I suppose those people that've loved their time in the sun get a few more days to keep going at that."
Lawes to captain England in Dublin
Courtney Lawes will captain the England side in Farrell's absence, with George Ford starting at fly-half and named a vice-captain.
Ford's start at fly-half is one of six changes to the side that defeated Wales 19-17 at Twickenham last Saturday.
In the front row, prop Ellis Genge is one of two changes to the forward pack, having won his 50th cap from the bench last Saturday, and he is joined by Will Stuart and Jamie George to complete the front row.
David Ribbans comes back into the side having successfully completed his return to play protocol and will be alongside Maro Itoje in the second row.
An unchanged back row sees captain Lawes continue with last weekend's player of the match Ben Earl and Billy Vunipola at No 8.
Ben Youngs and Ford make up the half-back partnership, with Manu Tuilagi starting his first game of the Summer Nations Series in the centres with Joe Marchant.
There is then a new set of wingers as Anthony Watson is named on the right wing with Elliot Daly on the left and Freddie Steward at full-back.
Among the replacements, Ollie Chessum is back in an England shirt for the first time since the Guinness Six Nations having recovered from an ankle injury, with Theo Dan, Joe Marler, Kyle Sinckler, Jack Willis, Danny Care, Marcus Smith and Ollie Lawrence completing the bench.
"It is a great challenge against the number one ranked team in the world and we are all excited for this fixture," added Borthwick.
"The game will be excellent preparation for us ahead of the Rugby World Cup."
England side to face Ireland
Starting XV: 15 Freddie Steward, 14 Anthony Watson, 13 Joe Marchant, 12 Manu Tuilagi, 11 Elliot Daly, 10 George Ford, 9 Ben Youngs; 1 Ellis Genge, 2 Jamie George, 3 Will Stuart, 4 Maro Itoje, 5 David Ribbans, 6. Courtney Lawes (C), 7 Ben Earl, 8 Billy Vunipola
Replacements: 16 Theo Dan, 17 Joe Marler, 18 Kyle Sinckler, 19 Ollie Chessum, 20 Jack Willis, 21 Danny Care, 22 Marcus Smith, 23 Ollie Lawrence
Ireland side to face England
Starting XV: 15 Hugo Keenan, 14 Mack Hansen, 13 Garry Ringrose, 12 Bundee Aki, 11 James Lowe, 10 Ross Byrne, 9 Jamison Gibson-Park, 1 Andrew Porter, 2 Dan Sheehan, 3 Tadhg Furlong, 4 Tadhg Beirne, 5 James Ryan (C), 6 Peter O'Mahony, 7 Josh van der Flier, 8 Cian Prendergast
Replacements: 16 Rob Herring, 17 Jeremy Loughman, 18 Finlay Bealham,19 Joe McCarthy, 20 Caelan Doris, 21 Conor Murray, 22 Jack Crowley, 23 Keith Earls