League bosses confirmed the game's worst-kept secret on Tuesday, with Penrith, Cronulla, Canberra and the Warriors bound for Vegas next year.
The matches will be two of four played at Allegiant Stadium on March 1, with Wigan and Warrington to kick off the event in a regular-season Super League fixture.
As revealed by AAP last week, Australia's women will also take on England in an international Test match on the same day at the venue.
There had initially been some thought given to playing four matches over a weekend-long festival of rugby league in the US.
But officials have instead opted for a full-day event, with kick-off to the Super League match at 1.30pm local time.
It means the action will begin at 8.30am in Australia, before the last NRL match between Penrith and Cronulla finishes at 5.30pm.
"This is the sequel. We're going to make sure this sequel is bigger and better," NRL CEO Andrew Abdo said.
"We have four fixtures, not two.
"The opportunity to showcase rugby league in Las Vegas, the sports and entertainment capital of America, in that stadium, it's a unique one.
"And the exciting thing is we're offering four new clubs, and ultimately their fan base and clubs the opportunity to experience what we all experienced in 2024."
The addition of a women's fixture is the most notable change, after officials had also weighed up whether to play an NRLW or women's State of Origin match.
The women's game is seen as the biggest area for growth for rugby league in the US, with no fully professional sports in the NFL or rugby in North America.
Officials also believe it can open a pipeline of talent up for NRLW clubs, once American athletes view the women's game.
"I don't think they would have seen female sport played at the level of speed, stamina, skill and intensity of rugby league football played by our female players," Abdo said.
"The opportunity (exists) to win new fans, and also the opportunity brings an enticing fixture for existing fans.
"It's just too good to pass on and we're excited by this fixture."
LAS VEGAS MATCHES ON MARCH 1
Wigan v Warrington (1.30pm PST / 8.30am AEDT)
Canberra v Warriors (4pm PST / 11am AEDT)
Australian Jillaroos v England Lionesses (6pm PST / 1pm AEDT)
Penrith v Cronulla (8.30pm PST / 3.30pm AEDT)
