Asked directly on Thursday whether Russia interfered in the election, Putin said: "Read my lips: No."
Putin's comments are the President's most emphatic denial of the accusations yet, and are the first he has directly made since US President Donald Trump took office in January.
Russian officials and spokespeople for Putin, however, have brushed off the claims several times as a political "witch hunt."
The denial comes as intelligence committees from both the House and Senate in the US investigate the alleged interference in the US election,
including possible ties between Russian officials
and members of Trump's campaign team.
The FBI is also conducting its own investigation.
Putin ridicules Kushner allegations
Putin also downplayed a meeting between Jared Kushner
-- Trump's son-in-law and close adviser -- and Sergey Gorkov, chairman of the Russian state development bank VneshEconomBank, which has been under US sanctions since July 2014.
Kushner is expected to give voluntary testimony to the Senate Intelligence Committee.
"Now there are some questions raised regarding the meetings with our bankers. But American bankers come to Russia and talk to us, don't they? Including our officials. How they can they work otherwise? Of course they hold meetings," Putin said, without explicitly naming the men in question.
"I think it's not in the interest of the American people to carry Russian-American relations to the point of absurdity, just to benefit the inner political agenda," Putin continued.
"What do we want? [To] break off the diplomatic relationship? Force the situation to the point of the 1960s [Cuban missile] crisis? And what's next?" Putin asked. "People who behave themselves so recklessly, what are they leading us to? Including the people of the USA. I think it's a big mistake and I hope the situation will normalize -- and the sooner the better."
The Russian leader also said that he would be open to meeting Trump on the side lines of an Arctic summit in Finland in September.
"If this happens, we will be glad to participate and I will be glad to take part in this event," he said.
Trump says he and Putin have never met, but both leaders have repeatedly said they are open to doing so.
Election meddling claims
The US government publicly announced in October that it was "confident" Russia orchestrated the hacking
of the Democratic National Committee (DNC) and other political organizations of the Democratic Party in the lead-up to the election.
And in January, days before Trump took office, the US intelligence community concluded that Putin had ordered an "influence campaign"
aimed at hurting Trump's rival, Democratic Party candidate Hillary Clinton.
The DNC hacks resulted in the public release of thousands of stolen emails, many of which included damaging revelations about the Democratic Party and Clinton's campaign.
Trump has flip-flopped on the issue, at first refusing to consider that Russia may have been behind the DNC hacks, but later conceding "I think it was Russia."
Kremlin spokesman Dmitry Peskov said in January that the charges against Russia "were not backed by anything" and were "made on a very amateur, emotional level."
"What we see is ... that all of this looks like is a full-scale witch hunt," Peskov said.
Before leaving office, former President Barack Obama ordered a full review into hacking aimed at influencing US elections going back to 2008.