Formation of a catalytically active dimer by tRNAVal -driven short ribozymes

A minizyme is a hammerhead ribozyme with a short oligonucleotide linker instead of stem/loop II. Minizymes with low activity as monomers form active dimeric structures with a common stem. We explored the use of dimeric minizymes as gene-inactivating agents by placing minizymes under the control of a tRNAVal promoter. The tRNA1Val portion of the transcript did not hinder dimerization as the tRNA-embed-ded minizyme formed an active dimeric structure. The cleavage activity of this minizyme that had been expressed either in vitro or in HeLa cells was almost one order of magnitude higher than that of the tRNAVal-embedded conventional hammerhead ribozyme. The tRNAVal-driven minizyme inhibited reporter gene activity (95%) whereas the tRNAVal-driven hammerhead ribozyme resulted in approximately 55% inhibition

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