SPECIFIC-INHIBITION OF DELTA-ANTIGEN BY IN-VITRO SYSTEM BY ANTISENSE OLIGODEOXYNUCLEOTIDE - IMPLICATIONS FOR TRANSLATION MECHANISM AND TREATMENT

Synthetic antisense oligodeoxynucleotides (ODNs) and a system containi ng transcription and translation coupled rabbit reticulocyte lysate we re used to develop a new model modulating the synthesis of small delta antigen which, in turn, inhibits the replication of HDV (hepatitis D virus). The ODN was stable for at least 50 min in this system al 37 de grees C. Unmodified 15-mer antisense D3 and D4, complementary to trans lation initiation region and coding region, respectively, inhibit the synthesis of small delta antigen by 95% at a concentration of 5 mu M, whereas antisenses complementary to 5' noncoding region, stop codon re gion and polyadenylation site were less effective. This system also sh owed a dose-dependent inhibitory effect of antisense D3 on the product ion of the target protein. However, the synthesis of E6 protein, an in ternal control, was not affected. These observations imply that this i n vitro system is convenient for rapid screening of effective antisens e compounds and offers a promising perspective for the investigation o f translation mechanisms and for the inhibition of HDV replication by antisense strategy. (C) 1997 Elsevier Science B.V.