Maria Corina Machado, a vocal supporter of anti-government protests that rocked the country earlier this year, vehemently rejected the charge as she left the attorney general's office after questioning.
"Today they have charged me with the crime of conspiracy," she said. "All the accusations and supposed evidence are false, and I reject them."
Under Venezuelan law she faces eight to 16 years in prison if convicted.
The attorney general's office yesterday said in a statement she had been charged with "conspiracy" for "allegedly having links to the assassination plan against the president."
Machado, 47, who says the evidence against her amounts to slander, was ousted from her seat in the National Assembly in March.
Along with Leopoldo Lopez, an opposition leader jailed since February on charges of inciting violent protests, she has been one of the most visible figures in an anti-government protest movement called "La Salida," or "The Exit."
The movement seeks to use street protests to press for Maduro's resignation.
The opposition and human rights groups have accused the government of using the judicial system to persecute dissidents.
Machado arrived at the attorney general's office accompanied by the leader of the main opposition coalition, Jesus Torrealba, Caracas mayor Antonio Ledezma and about 100 supporters.
Other opposition leaders rushed to her defense, including Henrique Capriles, the candidate who narrowly lost to Maduro in presidential elections last year.
Capriles dismissed the case against Machado as "a little circus put together by Nicolas (Maduro)."
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