Alice Albright, CEO of the Millennium Challenge Corporation, speaking at the ceremony called the signing historical between two "critical partners."
"Our partners want regional integration for trade, for economic growth, to increase the flow of goods across borders and to create more economic opportunities for their citizens," Albright said.
The signing of the deal, known as Benin-Niger Regional Transport Compact, will make available a grant of $504 million for connecting Benin's port of Cotonou to landlocked Niger's capital Niamey.
The deal will be further supported by contributions from Benin and Niger of $15 million each.
Benin President Patrice Tolon called the deal a major innovation in international cooperation between the U.S., Benin and Niger.
"If we increase our collaboration among African countries, countries that have small markets, this is a great way to increase the attractiveness of our countries," said Tolon.
"For a long time, we've considered this to be our natural port," Niger's President Mohamed Bazoum said speaking on Benin’s port of Cotonou
"We’ve developed all the necessary instruments to have relationships that make the economy of Niamey viable," said Bazoum.
He hailed the partnership with the United States and promised "institutional reforms" to support trade.
U.S. Secretary of State Antony Blinken also took part in the signing, which he said was key in building connectively within Africa, between African nations.
The signing of the regional compact deal was part of the U.S.-Africa Business Forum event hosted during day two of the U.S.-Africa Leaders Summit.
Some information for this report came from Agence-France Presse