Is your ribozyme design really correct?: A proposal of simple single turnover competition assay to evaluate ribozymes

Today, many nucleic acid enzymes are used in gene therapy and gene regulati ons. However, no simple assay methods to evaluate enzymatic activities, wit h which we judge the enzyme design, have been reported. Here, we propose a new simple competition assay for nucleic acid enzymes of different types to evaluate the cleaving efficiency of a target RNA molecule, of which the re cognition sites are different but overlapped. Two nucleic acid enzymes were added to one tube to make a competition of these two enzymes for one subst rate. The assay was used on two ribozymes, hammerhead ribozyme and hairpin ribozyme, and a DNA-enzyme. We found that this assay method is capable of a pplication to those enzymes, as a powerful tool for the selection and desig ning of RNA-cleaving enzymes.