Cell-free translation studies were carried out to compare the efficacy and specificity of four antisense structural types: DNA, phosphorothioate DNA (S-DNA), 2'-O-methyl RNA, and Morpholino oligos, a novel antisense structural type. In these studies, translational inhibition was assessed for two 20-mers of each structural type, where one 20-mer was complementary to its target sequence in rabbit alpha-globin mRNA and the other 20-mer contained three mispairs to that same target sequence. It is shown that at low concentration of antisense oligomer (50 nM) all four types provide high specificity, but the Morpholino oligos and 2'-O-methyl RNA afford better efficacy. At high oligomer concentration (3.5 microM), all four types provide high efficacy, but the Morpholino oligos and 2'-O-methyl RNA provide substantially better specificity than the DNA and S-DNA.