Trackhouse, which already competes in Nascar, will retain Miguel Oliveira and Raul Fernandez, who both competed with RNF this season, as its two riders.
"Joining the MotoGP world championship is a very exciting moment for our young company," said Trackhouse founder and owner Justin Marks in a statement.
"Trackhouse has worked from day one to recognize unique and compelling motorsport opportunities and being able to expand to a global series like MotoGP is a massive step in scaling the company."
Marks added that Trackhouse wanted to "help grow and amplify this amazing sport to millions of new fans in North America and beyond.”
The RNF team was excluded from the 2024 championship for "repeated infractions and breaches" affecting the competition's image, organisers said last week, the day after the final Grand Prix of the season, in Valencia.
RNF, founded by Malaysian Razlan Razali with the backing of state energy firm Petronas, entered MotoGP in 2019 under the name Sepang Racing Team.
After losing the financial support of Petronas last year, Razali took over the team and renamed it RNF. Romanian company CryptoDATA then became its majority shareholder at the end of 2022.
Razali had announced his departure on social media at the start of the weekend in Valencia.
The team, which used Yamaha bikes until 2022, was home to world champion Fabio Quartararo for his first two years in MotoGP.
The Frenchman then made the step up to Yamaha's factory team in 2021, trading places with Italian great Valentino Rossi who brought the curtain down on his career with the Malaysian team.
The 2024 MotoGP season kicks off on March 10th in Qatar.
