Selection of a ribozymes useful for, e.g. inhibiting gene expression and improving nuclease resistance

摘要

A method to select a ribozyme that can covalently modify the 2'-OH group of ribonucleic acids in trans is new. A method to select a ribozyme that can covalently modify the 2'-OH group of ribonucleic acids in trans, comprises: incubating a ribonucleic acid library with a reactant that can react with a ribonucleic acid 2'-OH group; selection and isolation of the ribonucleic acid that covalently binds to the reactant; amplifying the ribonucleic acid of step (b) to form cDNA; determining the sequence of the selected ribonucleic acid; cleavage of the selected ribonucleic acid in a substrate part (I) by a catalytic part (II = ribozyme); and testing whether the catalytically active part (II) can covalently modify the substrate part (I). Independent claims are also included for: a ribozyme obtained by a method as above; a DNA sequence that encodes a ribozyme as in (1); a vector containing DNA as in (2); a host cell containing a vector as in (3); and a method to produce a ribozyme as in (1).