The group that created the ad said it is "sorry" that Bush didn't want to be featured in the ad and will replace it with a new ad soon.
Former first lady Laura Bush is saying she wants to be removed from a pro-gay marriage ad created by the Respect for Marriage Coalition.
The ad features Bush speaking about gay marriage during a 2010 interview with Larry King on CNN.
"When couples are committed to each other and love each other, then they ought to have the same sort of rights that everyone has," she is shown as saying.
But Anne MacDonald, a spokeswoman for Bush, told the Dallas Morning News Wednesday that Bush wants the ad's creator to remove the clip.
"Mrs. Bush did not approve of her inclusion in this advertisement nor is she associated in any way with the group that made the ad," MacDonald said. "When she became aware of the advertisement Tuesday night, we requested that the group remove her from it."
Respect for Marriage Coalition said it is "sorry" that Bush didn't want to be featured in the ad, and will replace it with a new one this weekend, Politico reported.
"We used public comments for this ad from American leaders who have expressed support for civil marriage," the group said in a statement, according to Politico. "We appreciate Mrs. Bush's previous comments but are sorry she didn't want to be included in an ad. The ad launched a public education campaign that will now move to new and different voices that reflect the depth and breadth of our support."
The ad also features other prominent Republicans such as former Vice President Dick Cheney and former Secretary of State Colin Powell.
"Freedom means freedom for everyone," Cheney is shown saying in the ad. Cheney's daughter Mary married her partner, Heather Poe, in June.
Powell is shown saying, "Allowing them to live together with the protection of law, it seems to me is the way we should be moving in this country," a statement he made on CNN's "The Situation Room."
A spokeswoman for Powell said Thursday that they had not been contacted for permission to use the clip, CNN reported, but the spokeswoman said that Powell's "remarks are in the public domain and we have not asked them to remove it."
The ad also features President Barack Obama, who says, "Our journey is not complete until our gay brothers and sisters are treated like anyone else under the law," a statement he made during his inaugural address in January.
The Respect for Marriage Coalition launched a million-dollar television, print and online ad campaign for marriage equality Wednesday, saying that it featured bipartisan leaders including Obama, Cheney, Bush and Powell who were speaking out in support for gay and lesbian Americans.
The Supreme Court is expected to take up same-sex marriage this spring, with oral arguments scheduled for March 26 and 27.