The proposed sale of 316 RBS branches to Santander had collapsed after the Spanish banking giant pulled out of an agreed £1.65 billion purchase.
The UK state-owned bank had planned to sell the assets, which also included 40 banking centres for small and medium-sized businesses, in a deal approved by the European Commission.
RBS Group chief executive Stephen Hester said he was "disappointed" but promised there would be "no disruption" for customers and said the taxpayer-backed bank would now begin a process to seek a new buyer.
Mr Hester said: "I can assure all affected customers that there will be no disruption to the service they receive. It is business as usual in all of these branches, and customers don't need to take any action.
"It is of course disappointing that Santander decided to pull out of this transaction, especially for the customers and staff involved. However, RBS's strong progress in our restructuring plans means we can continue to provide a stable home for this business and its customers pending a further resolution.
"RBS will commence a new process of disposal and will provide a further update on this in due course."
Santander agreed to buy the assets in August 2010, broadly comprising the RBS branch business in England and Wales, and the NatWest branch business in Scotland, along with certain SME and corporate activities across the UK.
The deal was later approved by the European Commission after it investigated potential competition concerns.
The commission ordered RBS - 83% owned by the taxpayer - to sell the assets as a condition for receiving bailout aid from the government in the wake of the financial crisis.
An RBS statement tonight said: "Santander's decision follows extensive work by both parties to separate the business into a largely standalone form and to prepare the business, customers and staff for transfer.
"RBS is determined that the decision will have no impact on the service available to customers and will continue to work to fulfil its obligations to the European Commission."