Ribozyme mediated RNA double cleavage

The hairpin ribozyme is a small plant pathogenic RNA that achieves an active conformation by docking of its two helical domains in an antiparallel fashion. Our aim was the design of hairpin-like ribozymes with RNA double cleavage activity. We report on the construction of a reverse-joined hairpin ribozyme and the optimization of external parameters as well as structural properties for high cleavage activity. We show that under certain conditions the cleavage rate of the reverse-joined ribosome can be boosted to an area characteristic of the wild-type configured ribosome. Furthermore, the combination of the wild-type configured and the reverse-joined hairpin ribosome to a large ribosome with double cleavage activity is described.