Russian federal prison officials said Navalny, 47, collapsed while taking a walk on Friday and was unable to be resuscitated.
His death comes after years of activism against the Kremlin through his organization, the Anti-Corruption Foundation.
Biden and other leaders quickly blamed Putin for his death.
"What has happened to Navalny is yet more proof of Putin's brutality," Biden said. "No one should be fooled."
Navalny's wife, Yulia Navalnaya, also had harsh words for the Kremlin.
"They will pay for what they have done to our country, to our family, and my husband," she said to applause at the Munich Security Conference.
Navalny's death has also sparked a renewed debate around funding for Ukraine amid its war with Russia, with Congress deadlocked on passing new aid for Kyiv.
Republican presidential candidate Nikki Haley used the opportunity to blast former President Trump, who has at times praised Putin and has promised he will bring an end to the Ukraine war in 24 hours once in office.
"Putin did this," Haley wrote on X, formerly Twitter. "The same Putin who Donald Trump praises and defends."
Navalny was arrested in 2021 after he was poisoned by a Soviet nerve agent, which he pinned on Putin. The Kremlin denied involvement in the incident.
Navalny was sentenced to nine years in prison in 2022 on fraud charges and 19 years in prison last year on extremism charges. He had denied those charges.
Before his death, he was transferred to a remote Siberian prison colony in December.
In a documentary about his life, Navalny urged his supporters to never give up and suggested that his death would not mean the end of his cause.
"If they decided to kill me," he said, "then it means we are incredibly strong."
The Hill has published a timeline of Navalny's life.
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