Characterisation of catalytic nucleic acids targeting the survival of motor neuron messenger RNA

Catalytic nucleic acids have been used in a reverse genetics approach to study gene function. We are interested in applying this technology to study the autosomal recessive disease Spinal Muscular Atrophy (SMA) which is caused by loss of survival motor neuron gene (SMN) product leading to progressive motor neuron loss and muscular atrophy. Although the SMN gene is ubiquitously expressed, the cause for selective motor neuron loss is unknown. Embryonal lethality in mice has made it extremely difficult to generate animal models of SMA. We describe a procedure for selecting effective DNAzymes (DZ) and ribozymes (RZ) based on their ability to cleave the full length Smn mRNA at low magnesium concentrations, after a short time period, and at a low catalytic nucleic acid to target ratio. Using these criteria three effective RZ and DZ were generated. These results indicate that catalytic nucleic acids can effectively cleave Smn target RNA in an environment closely resembling that present in the cell and thus have potential for interference with Smn gene expression in cells and in vivo.