Helicase-attached novel hybrid ribozymes

Ribozymes have potential as therapeutic agents and in functional studies of genes of interest. The activities of ribozymes in vivo depend on the accessibility of ribozymes to a cleavage site in the target RNA. At present, the selection of a target site for ribozymes is often based on a computer-aided structural analysis of the target RNA or trial-and-error experiments in which vast numbers of ribozymes are tested systematically. To overcome this problem, we have engineered intracellularly produced ribozymes with unwinding activity in vivo. We found that attachment to ribozymes (hybrid ribozymes) of an RNA motif with the ability to interact with intracellular RNA helicases, which create hybrid ribozymes, enhances ribozyme activity significantly in vivo. Thus, hybrid ribozymes can catalyze cleavage at the specified target site within an RNA in vivo almost independently of the secondary or tertiary structure of the target RNA around the cleavage site.