Obama quipped that since his birth certificate had now been released, the businessman could focus "on the issues that matter. Like, did we fake the moon landing? What really happened in Roswell? And where are Biggie and 2Pac?"
He went on to deride Trump's starring role on The Celebrity Apprentice, joking that choosing who to fire would keep him awake at night.
Predicting what would happen if Trump's presidency bid was successful, Obama then showed a mocked-up image of the White House, which had been turned into a casino and golf course with girls in bikinis sitting outside in a hot tub.
Later on in the evening, comedian Seth Meyers also teased Trump, saying: "Donald Trump often talks about running as a Republican, which is surprising. I just assumed he was running as a joke."
Watch Obama's full speech below. He addresses Trump from 09.35: