The singer-songwriter will release 'Every Night I Say A Prayer' as a free download via Trax Records and 679 Artists, and as a limited-edition vinyl for Record Store Day on April 21.
Remixes by Joe Smooth and Tensnake are also included on the 12" version.
Little Boots performs at XOYO on Friday, May 5. Tickets are priced at £12.50 and are on sale now.
She will also mark the release of the single with an appearance at London's Rough Trade East on April 21.
Little Boots released the single 'Shake' - produced by Simian Mobile Disco's James Ford - last November.
She is currently working on the follow-up to her 2009 debut Hands.
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Watch the Little Boots 'Shake' music video below: