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In vitro selection of adenine‐dependent ribozyme against Tpl2/Cot oncogene

Hairpin ribozymes possess the properties of RNA sequence‐specific recognition and site‐specific cleavage. These properties make them a powerful extension of the antisense approach for the inhibition of gene expression. From a randomized RNA pool of hairpin ribozymes, using the systematic evolution of ligands by exponential enrichment, we have obtained an adenine‐dependent hairpin ribozyme, Tpl2/Cot (tumour progression locus 2) ribozyme, which cleaves the Tpl2/Cot kinase mRNA sequence at nucleotides A225↓G226 relative to the start codon of translation. This serine/threonine kinase activates the mitogen‐activated protein kinase pathway implicated in cell proliferation in cancer. The selected ‘Tpl2/Cot‐YL ribozyme’ efficiently cleaves its target sequence in cis and in trans; furthermore, the ribozyme efficiently cleaves a longer target sequence of 54 nucleotides in trans, as well as the full‐length mRNA.