Key result
Antisense oligonucleotides targeted at beta-globin mRNA inhibited both alpha- and beta-globin production in reticulocyte lysates, highlighting anomalies and the need for caution in antisense experiments.
In vitro experiments highlight anomalies in antisense control of translation, emphasizing caution in performing and interpreting antisense experiments.
Requires rigorous controls in antisense experiments to avoid artifacts; leaves open reliable translation to therapeutic gene targeting.
Experiments were carried out to better characterize antisense control of translation. Results in an E. coli system confirmed specific inhibition of poly(U) translation. At low concentrations, certain homopolymers (including poly(rA)) stimulated translation. Oligo(dA(n)) was inhibitory at n less than or equal to 8. Translation of globin mRNA in reticulocyte lysates indicated that ssDNA 15-mers targeted at beta-globin mRNA inhibited both alpha- and beta-globin production. Sequences targeted immediately downstream of the AUG were the least effective in inhibition. These and other anomalies are discussed here in relation to those of others, emphasizing caution in performing antisense experiments.
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Ricker et al. (1992) studied this question. Antisense oligonucleotides was evaluated on Inhibition of translation. Antisense oligonucleotides targeted at beta-globin mRNA inhibited both alpha- and beta-globin production in reticulocyte lysates, highlighting anomalies and the need for caution in antisense experiments.
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