Kinetic Characterization of Group II Intron Folding and Splicing

Group II introns are large self-splicing ribozymes found in bacterial genomes, in organelles of plants and fungi, and even in some animal organisms. Many organellar group II introns interrupt important housekeeping genes; therefore, their splicing is critical for the survival of the host organism. Group II introns are versatile catalytic RNAs: they facilitate their own excision from a pre-mRNA, they promote ligation of exons to form a translation-competent mature mRNA; they can act like mobile genomic elements and insert themselves into RNA and DNA targets with remarkable precision, which makes them attractive tools for genetic engineering. The first step in characterization of any group II intron is the evaluation of its catalytic activity and its ability to properly fold into the native functionally active structure. This chapter describes kinetic assays used to characterize folding and catalytic properties of group II intron-derived ribozymes.