Generation of stable mRNA fragments and translation of N-truncated proteins induced by antisense oligodeoxynucleotides

Binding of phosphorothioate-modified antisense oligodeoxynucleotides (AS ODNs) to target mRNAs is commonly thought to mediate RNA degradation or block of translation. Here we demonstrate cleavage of target mRNAs within the AS ODN binding region with subsequent degradation of the 5′ but not the 3′ cleavage product. Some, if not all, 3′ mRNA fragments lacked a 5′ cap structure, whereas their poly(A) tail length remained unchanged. Furthermore, they were efficiently translated into N-terminally truncated proteins as demonstrated in three settings: production of shortened hepadnaviral surface proteins, alteration of the subcellular localization of a fluorescent protein, and shift of the transcription factor C/EBPα isoform expression levels. Thus, AS treatment may result in the synthesis of N-truncated proteins with biologically relevant effects.