Sequence-specific inhibition of human immunodeficiency virus (HIV) reverse transcription by antisense oligonucleotides: comparative study in cell-free assays and in HIV-infected cells
| Autor: | Claude Hélène, Genevieve Degols, Simon Litvak, Maritta Perala-Heape, Bernard Lebleu, Leila Sarih-Cottin, Bruno Bordier |
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| Rok vydání: | 1995 |
| Předmět: |
DNA Replication
RNase P Molecular Sequence Data Ribonuclease H Polymerase Chain Reaction Virus Cell Line Proviruses Complementary DNA Humans Polylysine RNase H DNA Primers Multidisciplinary biology Base Sequence Cell-Free System Chemistry Oligonucleotide DNA replication RNA-Directed DNA Polymerase Oligonucleotides Antisense Virology Molecular biology Reverse transcriptase HIV Reverse Transcriptase Kinetics biology.protein HIV-1 RNA Viral Reverse Transcriptase Inhibitors Primer binding site Research Article |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 92(20) |
| ISSN: | 0027-8424 |
| Popis: | We have investigated two regions of the viral RNA of human immunodeficiency virus type 1 (HIV-1) as potential targets for antisense oligonucleotides. An oligodeoxynucleotide targeted to the U5 region of the viral genome was shown to block the elongation of cDNA synthesized by HIV-1 reverse transcriptase in vitro. This arrest of reverse transcription was independent of the presence of RNase H activity associated with the reverse transcriptase enzyme. A second oligodeoxynucleotide targeted to a site adjacent to the primer binding site inhibited reverse transcription in an RNase H-dependent manner. These two oligonucleotides were covalently linked to a poly(L-lysine) carrier and tested for their ability to inhibit HIV-1 infection in cell cultures. Both oligonucleotides inhibited virus production in a sequence- and dose-dependent manner. PCR analysis showed that they inhibited proviral DNA synthesis in infected cells. In contrast, an antisense oligonucleotide targeted to the tat sequence did not inhibit proviral DNA synthesis but inhibited viral production at a later step of virus development. These experiments show that antisense oligonucleotides targeted to two regions of HIV-1 viral RNA can inhibit the first step of viral infection--i.e., reverse transcription--and prevent the synthesis of proviral DNA in cell cultures. |
| Databáze: | OpenAIRE |
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