Design and Experimental Evolution of trans-Splicing Group I Intron Ribozymes.

Group I intron ribozymes occur naturally as -splicing ribozymes, in the form of introns that do not require the spliceosome for their removal. Instead, they catalyze two consecutive -phosphorylation reactions to remove themselves from a primary transcript, and join the two flanking exons. Designed, -splicing variants of these ribozymes replace the 3'-portion of a substrate with the ribozyme's 3'-exon, replace the 5'-portion with the ribozyme's 5'-exon, or insert/remove an internal sequence of the substrate. Two of these designs have been evolved experimentally in cells, leading to variants of group I intron ribozymes that splice more efficiently, recruit a cellular protein to modify the substrate's gene expression, or elucidate evolutionary pathways of ribozymes in cells. Some of the artificial, -splicing ribozymes are promising as tools in therapy, and as model systems for RNA evolution in cells. This review provides an overview of the different types of -splicing group I intron ribozymes that have been generated, and the experimental evolution systems that have been used to improve them.