Greek airline Minoan Air will operate twice-daily flights on Mondays, Tuesdays and Thursdays from March, increasing frequency to six days a week from May 10. The service has been pitched at giving business passengers a "practical choice".
The fastest rail journey is five hours and 22 minutes – with some trips taking more than six hours – compared to an hour-and-a-half in the air.
Flights between Edinburgh Airport and London Oxford Airport were launched in March 2010 by Varsity Express but stopped after only six days.
Andi Pargeter, managing director of London Oxford Airport, said: "Minoan flights to Edinburgh will take an hour-and-a-half, making a day return to Scotland a practical choice.
"The two cities have a lot of synergy, but rail journeys between Oxford and Edinburgh can typically take six hours or more, and with rail fares between the two cities averaging more than £200, the direct flights will definitely appeal to business passengers who have previously had to factor an overnight stay into their visits."