Greene obtained degrees at both Cambridge and Oxford. He then went to London, where he became an intimate of its underworld. He wrote more than 35 works between 1580 and 1592. To be certain of supplying material attractive to the public, Greene at first slavishly followed literary fashions. His first model was John Lyly's Euphues.

In the later 1580s Greene wrote prose pastorals in the manner of Sir Philip Sidney's Arcadia, interspersed with charming, often irrelevant lyrics that have given Greene a reputation as a poet. The best of his pastorals is Pandosto (1588), the direct source of Shakespeare's (Shakespeare, William) The Winter's Tale.

About 1590 Greene began to compose serious didactic works. Beginning with Greenes never too late (1590), he related prodigal son stories. That Greene drew on his own experience is evident from the tract Greenes groats-worth of witte, bought with a million of repentance, printed posthumously in 1592 with Greene's admission that Roberto's experiences were essentially his own. In Groats-worth appears the first printed reference to Shakespeare, assailed as “an upstart Crow, beautified with our feathers, that with his Tygers heart wrapt in a Players hide, supposes he is as well able to bumbast out a blanke verse as the best of you . . . in his owne conceit the onely Shake-scene in a countrie.” (The words in italics are from Shakespeare's I Henry VI.) Greene is thought to be criticizing Shakespeare the actor.

Greene's writings for the theatre present numerous problems; the dating of his plays is conjectural, and his role as collaborator has produced much inconclusive discussion. With The Honorable Historie of frier Bacon, and frier Bongay (written c. 1591, published 1594), the first successful romantic comedy in English, Greene realized his comic talent in drama. In The Scottish Historie of James the fourth, slaine at Flodden (written c. 1590, published 1598) he used an Italian tale but drew on fairy lore for the characters of Oberon and Bohan. It was a forerunner of As You Like It and A Midsummer Night's Dream. As Marlowe anticipated the tragedies of Shakespeare, so, in a lesser way, Greene furnished him a model in dramatic comedy and romance.

In his last year Greene wrote exposés of the Elizabethan underworld, such as A Notable Discovery of Coosnage (1591) and the successful and amusing A disputation betweene a hee conny-catcher and a shee conny-catcher (1592).