Development of Artificial Ribonucleases

We have developed Oligonucleotide Based Artificial Nucleases (OBANs) with the aim to produce artificial enzymes that can cleave the messenger-RNA sequences arising from the M-BCR/ABL gene or from other genetic or viral diseases. The goal is to utilize the PNAzyme platform (Appendix 1.) and from this develop artificial nucleases that can be used as tools for in vitro cell assays and eventually for in vivo gene silencing. The currently developed system does still do not quite give sufficient cleavage rates (an increase of about one order of magnitude is needed) for gene silencing applications and the binding constant (metal to chelate) should be improved to avoid partial dissociation of the metal ion. Alternatively, the system can be altered to make use of a non-toxic metal ion (e.g., Mg2+) that is abundant in the biological system.

The PNAzymes are at the moment in a very exciting state of the development. These highly site- and sequence-selective PNAzyme systems give cleavage rates higher than any other systems reported in the literature so far. Student will synthesize and evaluate new PNAzymes by RP-HPLC/IE-HPLC/MS/UV methodologies.