Originally, an artist named Akema Johnson-Day was slated to provide lead vocals for the song with Virgo Williams on background vocals, however when a scheduling conflict prevented Akema from taking part in the production, Virgo Williams's vocals became the lead vocals. Lil Jon was an A&R Director for So So Def at the time. Rodney Terry and Carl Mahone produced it, and it was the second version of the beat currently used in the song.
The music video features a pool party. The group does not appear in the video, but Jermaine Dupri does.
Twenty years after the song's original chart run, a resurgence in its popularity driven by the Running Man Challenge viral meme prompted its re-entry on the charts dated May 21, 2016. On the strength of 35,000 digital download sales and 12.3 million domestic streams, "My Boo" re-entered the R&B chart, which had long since been renamed Hot R&B/Hip-Hop Songs, at number 14, and the Hot 100 at number 29, surpassing the single's initial peaks on both charts. It subsequently peaked at numbers 10 and 27, respectively.
In 2016, twenty years after its initial release, "My Boo" gained notoriety through the Running Man Challenge, a viral meme in which participants filmed and shared short clips of themselves performing a dance resembling running to the song. The original videos were posted on Vine by high school students Kevin Vincent and Jeremiah Hall in Hillside, New Jersey. Inspired by Vincent and Hall's video, two players from the University of Maryland basketball program, Jared Nickens and Jaylen Brantley, filmed their own versions of the Running Man Challenge, spreading the meme.
The challenge went viral on Vine, and Vincent, Hall, Nickens and Brantley were invited to perform it on The Ellen DeGeneres Show. Many other athletes from various sports, a number of celebrities and the general public embraced the trend and produced their own takes of the dance.
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