Brennan said that such meddling "would destroy any near-term prospect of improvement" in relations between the United States and Russia. The FSB chief, Alexander Bortnikov, twice denied that Russia was waging such a campaign but said he would carry Brennan's message to Russian President Vladimir Putin.
"I believe I was the first US official to brace Russia on this matter," Brennan said during his first appearance before the House Intelligence Committee as part of that panel's investigation of a Russian influence campaign in the 2016 presidential election.
Brennan was among the top officials who briefed then-President-elect Donald Trump on Russia's goals - which represented the consensus view of the CIA, the FBI and the Office of the Director of National Intelligence.
Brennan became so alarmed by the Russian intervention last northern autumn that he held classified meetings with top congressional officials to impress upon them the unprecedented nature of Moscow's interference.
Brennan testified that he was disturbed by intelligence that surfaced last year showing a pattern of contacts between Russian agents or representatives and people with links to the Trump campaign.
"That raised concerns in my mind," Brennan said. He emphasised that the information he saw did not amount to proof of collusion or cooperation between Trump associates and Russia, but he said that it "served as the basis for the FBI investigation".
With that remark, Brennan appeared to identify the point of origin of the FBI investigation that began in July - the first time a US official has provided insight into what prompted the bureau probe.
Brennan said that the targets of those Russian approaches may not have been aware of the nature of the contacts because Russian services often disguise their efforts by using intermediaries.
"Many times, [US individuals] do not know that the individual they are interacting with is a Russian," Brennan said. Russians use an array of espionage tools, including blackmail over compromising information, to coerce treason from US officials who "do not even realise they are on that path until it gets too late".
Brennan declined to name any US individuals, but that remark appeared to be in reference to former National Security Adviser Michael Flynn, who was forced to resign after misleading statements about his contacts with the Russian Ambassador.
Brennan was also asked about Trump's disclosure of highly classified information to the Russian Foreign Minister and Ambassador in a White House meeting this month. Brennan said that the CIA at times provided tips about terrorist plots to the Kremlin, but he indicated that Trump violated key protocols.
Sensitive information should only be passed through intelligence services, not divulged to foreign ministers or ambassadors, Brennan said. "Neither did it go in the proper channels nor did the originating agency have the opportunity to clear language for it."
The former CIA chief is the latest senior Obama Administration official to appear publicly before Congress in hearings that have often produced damaging headlines for Trump.
Earlier this month, former acting Attorney-General Sally Yates testified that she expected White House officials to "take action" after warning that Flynn had misled Administration officials about his contacts with Russia.
At that same hearing, former director of national intelligence James Clapper said that Moscow's leaders "must be congratulating themselves for having exceeded their wildest expectations with a minimal expenditure of resource," a reference not only to the outcome of the 2016 race but also to the chaos that has characterised the early months of the Trump Administration.
Brennan has feuded publicly with Trump over the President's treatment of intelligence agencies. In January, he lashed out at Trump for comparing US spy agencies to Nazi secret police.
Brennan was particularly offended by Trump's remarks during a speech at CIA headquarters on the day after he was inaugurated. Trump used the CIA's Memorial Wall - a collection of engraved stars marking the lives of agency operatives killed in the line of duty - to launch a rambling speech in which he bragged about his election victory.
Brennan called the appearance "despicable" and said that Trump should be "ashamed."