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The FAA has previously strongly advised U.S. operators against flying over Syria. The new rule requires them to contact the agency before operating in the airspace.
"Due to the presence of anti-aircraft weapons among the extremist groups and ongoing fighting in various locations throughout Syria, there is a continuing significant potential threat to civil aviation operating in Syrian airspace," the FAA said in a statement.
The agency said it is prudent to prohibit U.S. carriers from flying into, out of, and over Syria, based on an "updated assessment of the risk" and the "lack of any requests from operators wishing to fly in this airspace".
Armed extremist groups in Syria are known to be equipped with a variety of anti-aircraft weapons which could threaten civilian aircraft, said the statement, adding that the opposition groups had successfully shot down Syrian military aircraft using these weapon systems during the course of the conflict.