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 containing transcription and translation coupled rabbit reticulocyte lysate were 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 at 37 degrees C. Unmodified 15-mer antisense D3 and D4, complementary to translation initiation region and coding region, respectively, inhibit the synthesis of small delta antigen by 95% at a concentration of 5 microM, whereas antisenses complementary to 5' noncoding region, stop codon region and polyadenylation site were less effective. This system also showed a dose-dependent inhibitory effect of antisense D3 on the production of the target protein. However, the synthesis of E6 protein, an internal control, was not affected. These observations imply that this in vitro system is convenient for rapid screening of effective antisense compounds and offers a promising perspective for the investigation of translation mechanisms and for the inhibition of HDV replication by antisense strategy.
Chen
T Z
TZ
Department of Medicine, Veterans General Hospital-Taipei, Taiwan, R.O.C.
Wu
J C
JC
Au
L C
LC
Choo
K B
KB
D000818:Animals; D002474:Cell-Free System; D003673:Defective Viruses; D004305:Dose-Response Relationship, Drug; D004355:Drug Stability; D018963:Hepatitis Antigens / Q000187:drug effects* / Q000378:metabolism*; D003698:Hepatitis Delta Virus / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D035923:Hepatitis delta Antigens; D016376:Oligonucleotides, Antisense / Q000378:metabolism / Q000494:pharmacology*; D014176:Protein Biosynthesis / Q000187:drug effects*; D016601:RNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D011817:Rabbits; D012156:Reticulocytes / Q000737:chemistry / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects
D018963:Hepatitis Antigens; D035923:Hepatitis delta Antigens; D016376:Oligonucleotides, Antisense; D016601:RNA-Binding Proteins; C442244:hepatitis delta virus large antigen