The sCYMV1 hairpin ribozyme: targeting rules and cleavage of heterologous RNA

Abstract

The catalytic center of the RNA from the negative strand A*GUA sequence where * is the site of cleavage. When of the satellite RNA of chicory yellow mottle virus type 1 each nucleotide in this sequence was changed to each of (sCYMV1) is in the hairpin ribozyme family, has catalytic the other three nucleotides, catalytic activity decreased 66-activity, and cleaves substrates before a preferred GUA 100%. RNA targets, containing this A*GUA sequence, sequence. This is different from that of the satellite RNA were located in both human papillomavirus and HIV-1. from the negative strand of tobacco ringspot virus (sTRSV)Ribozymes were developed which efficiently cleaved these which prefers a GUC sequence at the site of cleavage. The targets in vitro. These results identify a new class of hairpin sCYMV1 hairpin ribozyme has now been developed for ribozymes capable of cleaving substrates before a precleaving heterologous RNA substrates. When helix 1 was ferred GUA sequence rather than the GUC preferred by extended from the native 5 bp to 6 bp with a newly added the sTRSV hairpin ribozyme. This expands the repertoire A:U base pair, catalytic activity increased three-fold. The of target sites available for gene therapy using the hairpreferred sequence for the substrate loop was the native pin ribozyme.

A comparative survey of the influence of small self-cleaving ribozymes on gene expression in human cell culture

“…The hairpin ribozyme ( Figure 1(C) ) belongs to one of the earliest described ribozyme classes. Here, a sequence derived from the chicory yellow mottle virus (sCYMV1) was used [ 65 ]. Although every catalysed reaction is an equilibrium between cleavage and re-ligation, the equilibrium of most known ribozymes is predominantly found on the side of the cleavage products.…”