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"Accusing the Islamic Republic of seeking nuclear arms is a blatant lie. We are fundamentally opposed to nuclear weapons," the English-language Press TV quoted Khamenei as saying.
"We consider the use and proliferation of nuclear arms as forbidden," he added.
The Iranian supreme leader said the United States and its Western allies are promoting the policy of Islamophobia, in a bid to prevent the Islamic world from maintaining its unity, Press TV reported.
"The U.S. authorities, who consider Iran's missiles as threats and claim Tehran is producing nuclear bombs, are well aware that their accusations are baseless and contrary to the fact," Khamenei said.
He insisted that Iran would resist such a policy and would never step back in the face of any aggression.
The United States and other Western countries claim that Iran intends to secretly develop nuclear weapons. The UN Security Council also requires Iran to suspend its uranium enrichment activity.
Iran, however, insists that its nuclear plan is only for peaceful purposes, vowing to continue its uranium enrichment activity despite pressure and sanctions from Western countries.