The evaluation of a hairpin ribozyme to down regulate maize alcohol dehydrogenase expression in vivo
Catalytic RNA is being used as a new tool to study the down regulation of gene expression. Our objective was to develop a model system to test the ability of the "hairpin" ribozyme to cleave target message RNA in vivo. A synthetic hairpin ribozyme was designed to cleave the maize alcohol dehydrogenase 1 (Adh1) message. Adh1 was chosen because activity of the protein is easily assayable and a selection system for stable transformants is available. The synthetic ribozyme (RADH) was designed to target native Adh1 mRNA at nucleotide position 3093 containing the requisite GUC cleavage site. Two pUC19 based vectors were engineered for in vivo expression of RADH at high levels in maize protoplasts. Transient and stable expression assays were performed to evaluate cleavage of the Adh1 mRNA transcript.