The 31-year-old from Coleraine was a silver medallist, along with his brother Peter, in the lightweight men's four at the 2012 Olympics in London.
In his new position of assistant coach, Chambers will help prepare Cambridge for their annual race against Oxford.
Chambers won his world titles in 2007 and 2010 in the lightweight men's four.
He won European silvers in 2014 and 2015 in the lightweight men's double scull, before taking world silver in 2015 with his 2016 Olympic partner Will Fletcher.
At this summer's Rio Games, Chambers and Fletcher missed out on a place in the final as they finished fourth in their semi-final.
"I have had a career in rowing which has been much greater than I could have ever dreamed.  I never believed that I would make one Olympic Games never mind, three," said Chambers.
"I have always been proud to represent my country as a lightweight at rowing, and I have enjoyed my career through all the ups and downs."
Two-times world champion Richard Chambers has retired from rowing to take up a coaching role with Cambridge University Boat Club.