Specific inhibition of human immunodeficiency virus type 1 replication by RNA transcribed in sense and antisense orientation from the 5'-leader/gag region.
The inhibitory effects of expression plasmids on HIV-1 replication were studied in a transient assay system. Test plasmids were co-microinjected with non-defective proviral HIV-1 DNA into a colon-carcinoma cell line (SW480) and the resulting infectious HIV-1 was quantitated after amplification in cocultivated CD4+ MT-4 cells. At a molar ratio of 1:1 and 5:1 plasmids capable of expressing a 410 bp HIV-1 fragment as antisense or sense transcript respectively both specifically inhibited HIV-1 replication up to 70%. This effect was specific for HIV-1 sequences and was not observed upon expression of unrelated RNA-segments. At a molar excess equal to or greater than 15:1, additional inhibitory effects were seen with control plasmids carrying only the strong human cytomegalovirus immediate early (HCMV IE) promoter/enhancer element. The reasons for these findings are discussed.
Institut für Virusforschung, Deutsches Krebsforschungszentrum, Heidelberg, F.R.G.
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G
Sczakiel
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Pawlita
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A
Kleinheinz
D015704:CD4 Antigens / Q000032:analysis; D002460:Cell Line; D003110:Colonic Neoplasms; D004279:DNA, Viral / Q000008:administration & dosage / Q000235:genetics; D005784:Gene Amplification; D015683:Gene Products, gag / Q000235:genetics*; D005814:Genes, Viral; D015488:HIV Antigens / Q000032:analysis; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008845:Microinjections; D010957:Plasmids; D021382:Protein Sorting Signals / Q000235:genetics*; D011533:Proviruses / Q000235:genetics; D012313:RNA / Q000235:genetics*; D016372:RNA, Antisense; D012333:RNA, Messenger / Q000037:antagonists & inhibitors*; D014158:Transcription, Genetic; D014779:Virus Replication
D015704:CD4 Antigens; D004279:DNA, Viral; D015683:Gene Products, gag; D015488:HIV Antigens; D021382:Protein Sorting Signals; D016372:RNA, Antisense; D012333:RNA, Messenger; D012313:RNA