| Summary: | <p>Abstract</p> <p>Background</p> <p>Allosteric ribozymes (aptazymes) that have extraordinary activation parameters have been generated <it>in vitro </it>by design and selection. For example, hammerhead and ligase ribozymes that are activated by small organic effectors and protein effectors have been selected from random sequence pools appended to extant ribozymes. Many ribozymes, especially self-splicing introns, are known control gene regulation or viral replication <it>in vivo</it>. We attempted to generate Group I self-splicing introns that were activated by a small organic effector, theophylline, and to show that such Group I aptazymes could mediate theophylline-dependent splicing <it>in vivo</it>.</p> <p>Results</p> <p>By appending aptamers to the Group I self-splicing intron, we have generated a Group I aptazyme whose <it>in vivo </it>splicing is controlled by exogenously added small molecules. Substantial differences in gene regulation could be observed with compounds that differed by as little as a single methyl group. The effector-specificity of the Group I aptazyme could be rationally engineered for new effector molecules.</p> <p>Conclusion</p> <p>Group I aptazymes may find applications as genetic regulatory switches for generating conditional knockouts at the level of mRNA or for developing economically viable gene therapies.</p>
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