THE 5' CAP: A NOVEL TARGET FOR ANTISENSE TECHNOLOGY

THE MAJORITY OF EUKARYOTIC AND VIRAL SMALL NUCLEAR AND MESSENGER RNAS ANALYZED TO DATE HAVE A UNIQUE CHEMICAL STRUCTURE AT THEIR 5' TERMINUS THAT IS REQUIRED IN VARYING DEGREES FOR THEIR PROCESSING, TRANSPORT TO THE CYTOPLASM, STABILITY, AND EFFICACY IN TRANSLATION. THIS RESEARCH WILL DETERMINE IF THE REMOVAL OR ALTERATION OF THE 5' CAP OF MESSENGER RNA BY A REACTIVE MOIETY COUPLED TO AN ANTISENSE OLIGONUCLEOTIDE IS A VIABLE APPROACH FOR THE REGULATION OF GENE EXPRESSION. IT IS PLANNED FIRST TO SCREEN FOR REACTIVE FUNCTIONAL GROUPSTHAT CATALYTICALLY CLEAVE THE PHOSPHOANHYDRIDE, PHOSPHOMONOESTER ANHYDRIDE, OR N-GLYCOSIDIC BOND OR REACT WITH THE ELECTROPHILIC CARBON AT THE EIGHT POSITION OF THE METHYLATED GUANINE RESIDUE. REACTIVE GROUPS FROM THIS SCREENING PROCESS WILL THEN BE TETHERED TO AN APPROPRIATE ANTISENSE OLIGONUCLEOTIDE. MOLECULAR MECHANICS AND MODELING USING COMPUTER GRAPHICS SOFTWARE PACKAGES WILL BE USED TO ASSIST IN THE DESIGN OF THE ANTISENSE HYBRID, SUCH AS IN THE TETHER LENGTH AND COMPOSITION. PHASE I CLEAVAGE ASSAYS WILL BE DONE BY FPLC USING CAPPED MONORIBONUCLEOTIDES AS SUBSTRATE.