The sisters were supposed to compete in that event at Roland Garros for the first time since 2010 after receiving a wild card from the tournament.
But they withdrew Wednesday. No reason was given by the tournament.
Venus said she was bothered by pain in her back during a three-set loss in singles Sunday. Serena is into the third round of singles.
They have won a total of 13 Grand Slam trophies in women's doubles, including at the French Open in 1999 and 2010.