Comparative inhibition of ras p21 protein synthesis with phosphorus-modified antisense oligonucleotides

A rabbit reticulocyte lysate translation assay was used to quantitatively compare a series of antisense oligodeoxyribonucleotides (11-mers) having different internucleoside linkages and various degrees of complementarity (100-80%) with the start codon and downstream 8 bases of Balb-ras p21 mRNA. The oligomers had either contiguous phosphodiester, or alternating methylphosphonate-phosphodiester, or contiguous methylphosphonate, or contiguous phosphorothioate linkages. Under the conditions used for the assay, all of the test compounds when present in about 10(3)-10(4) excess over mRNA (15 nM mRNA) inhibited protein synthesis to a degree which was dependent on both the concentration and sequence of the oligomer. At low concentrations (12.5-25 microM), the phosphorothioate analogs were the most potent inhibitors of p21 protein synthesis; however, a sequence non-specific effect for these oligomers was dominant at higher concentrations of oligomer (100-200 microM). The methylphosphonate oligomers appeared to be slightly more discriminant. Relative hybridization strengths were assessed by melting (Tm) studies using a DNA oligomer target to mimic the mRNA.