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. 1992 Sep;66(9):5576–5581. doi: 10.1128/jvi.66.9.5576-5581.1992

Tat- and Rev-directed antisense RNA expression inhibits and abolishes replication of human immunodeficiency virus type 1: a temporal analysis.

G Sczakiel 1, M Oppenländer 1, K Rittner 1, M Pawlita 1
PMCID: PMC289117  PMID: 1501292

Abstract

Replication of human immunodeficiency virus type 1 (HIV-1) was inhibited by stable intracellular expression of antisense RNA in the human T-lymphoid cell line Jurkat. When the viral subregion encoding the HIV-1 activator proteins was targeted, the extent of antisense RNA-mediated inhibition was greater than 97% during the first 2 weeks postinfection. Later in the time course, productive HIV-1 infection broke through at high initial infective doses. However, at initial multiplicities of infection equal to or smaller than 0.1, HIV-1 production was not detectable during the 5 weeks of observation. The results underline the effectiveness of stable intracellular antisense RNA expression in inhibiting HIV-1 replication.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Agrawal S., Ikeuchi T., Sun D., Sarin P. S., Konopka A., Maizel J., Zamecnik P. C. Inhibition of human immunodeficiency virus in early infected and chronically infected cells by antisense oligodeoxynucleotides and their phosphorothioate analogues. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7790–7794. doi: 10.1073/pnas.86.20.7790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chang L. J., Stoltzfus C. M. Inhibition of Rous sarcoma virus replication by antisense RNA. J Virol. 1987 Mar;61(3):921–924. doi: 10.1128/jvi.61.3.921-924.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Coleman J., Hirashima A., Inokuchi Y., Green P. J., Inouye M. A novel immune system against bacteriophage infection using complementary RNA (micRNA). Nature. 1985 Jun 13;315(6020):601–603. doi: 10.1038/315601a0. [DOI] [PubMed] [Google Scholar]
  4. Cotten M., Schaffner G., Birnstiel M. L. Ribozyme, antisense RNA, and antisense DNA inhibition of U7 small nuclear ribonucleoprotein-mediated histone pre-mRNA processing in vitro. Mol Cell Biol. 1989 Oct;9(10):4479–4487. doi: 10.1128/mcb.9.10.4479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dropulić B., Lin N. H., Martin M. A., Jeang K. T. Functional characterization of a U5 ribozyme: intracellular suppression of human immunodeficiency virus type 1 expression. J Virol. 1992 Mar;66(3):1432–1441. doi: 10.1128/jvi.66.3.1432-1441.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ensoli B., Barillari G., Salahuddin S. Z., Gallo R. C., Wong-Staal F. Tat protein of HIV-1 stimulates growth of cells derived from Kaposi's sarcoma lesions of AIDS patients. Nature. 1990 May 3;345(6270):84–86. doi: 10.1038/345084a0. [DOI] [PubMed] [Google Scholar]
  7. Frankel A. D., Pabo C. O. Cellular uptake of the tat protein from human immunodeficiency virus. Cell. 1988 Dec 23;55(6):1189–1193. doi: 10.1016/0092-8674(88)90263-2. [DOI] [PubMed] [Google Scholar]
  8. Freier S. M., Kierzek R., Jaeger J. A., Sugimoto N., Caruthers M. H., Neilson T., Turner D. H. Improved free-energy parameters for predictions of RNA duplex stability. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9373–9377. doi: 10.1073/pnas.83.24.9373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Friedmann T. Progress toward human gene therapy. Science. 1989 Jun 16;244(4910):1275–1281. doi: 10.1126/science.2660259. [DOI] [PubMed] [Google Scholar]
  10. Goodchild J., Agrawal S., Civeira M. P., Sarin P. S., Sun D., Zamecnik P. C. Inhibition of human immunodeficiency virus replication by antisense oligodeoxynucleotides. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5507–5511. doi: 10.1073/pnas.85.15.5507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Harada S., Koyanagi Y., Yamamoto N. Infection of HTLV-III/LAV in HTLV-I-carrying cells MT-2 and MT-4 and application in a plaque assay. Science. 1985 Aug 9;229(4713):563–566. doi: 10.1126/science.2992081. [DOI] [PubMed] [Google Scholar]
  12. Hélène C., Toulmé J. J. Specific regulation of gene expression by antisense, sense and antigene nucleic acids. Biochim Biophys Acta. 1990 Jun 21;1049(2):99–125. doi: 10.1016/0167-4781(90)90031-v. [DOI] [PubMed] [Google Scholar]
  13. Joshi S., Van Brunschot A., Asad S., van der Elst I., Read S. E., Bernstein A. Inhibition of human immunodeficiency virus type 1 multiplication by antisense and sense RNA expression. J Virol. 1991 Oct;65(10):5524–5530. doi: 10.1128/jvi.65.10.5524-5530.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kim S. Y., Byrn R., Groopman J., Baltimore D. Temporal aspects of DNA and RNA synthesis during human immunodeficiency virus infection: evidence for differential gene expression. J Virol. 1989 Sep;63(9):3708–3713. doi: 10.1128/jvi.63.9.3708-3713.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Larder B. A., Darby G., Richman D. D. HIV with reduced sensitivity to zidovudine (AZT) isolated during prolonged therapy. Science. 1989 Mar 31;243(4899):1731–1734. doi: 10.1126/science.2467383. [DOI] [PubMed] [Google Scholar]
  16. Larder B. A., Kemp S. D. Multiple mutations in HIV-1 reverse transcriptase confer high-level resistance to zidovudine (AZT). Science. 1989 Dec 1;246(4934):1155–1158. doi: 10.1126/science.2479983. [DOI] [PubMed] [Google Scholar]
  17. Maher L. J., 3rd, Dolnick B. J. Comparative hybrid arrest by tandem antisense oligodeoxyribonucleotides or oligodeoxyribonucleoside methylphosphonates in a cell-free system. Nucleic Acids Res. 1988 Apr 25;16(8):3341–3358. doi: 10.1093/nar/16.8.3341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Malim M. H., Böhnlein S., Hauber J., Cullen B. R. Functional dissection of the HIV-1 Rev trans-activator--derivation of a trans-dominant repressor of Rev function. Cell. 1989 Jul 14;58(1):205–214. doi: 10.1016/0092-8674(89)90416-9. [DOI] [PubMed] [Google Scholar]
  19. Mann D. A., Frankel A. D. Endocytosis and targeting of exogenous HIV-1 Tat protein. EMBO J. 1991 Jul;10(7):1733–1739. doi: 10.1002/j.1460-2075.1991.tb07697.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Matsukura M., Shinozuka K., Zon G., Mitsuya H., Reitz M., Cohen J. S., Broder S. Phosphorothioate analogs of oligodeoxynucleotides: inhibitors of replication and cytopathic effects of human immunodeficiency virus. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7706–7710. doi: 10.1073/pnas.84.21.7706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ratner L., Haseltine W., Patarca R., Livak K. J., Starcich B., Josephs S. F., Doran E. R., Rafalski J. A., Whitehorn E. A., Baumeister K. Complete nucleotide sequence of the AIDS virus, HTLV-III. Nature. 1985 Jan 24;313(6000):277–284. doi: 10.1038/313277a0. [DOI] [PubMed] [Google Scholar]
  22. Rhodes A., James W. Inhibition of human immunodeficiency virus replication in cell culture by endogenously synthesized antisense RNA. J Gen Virol. 1990 Sep;71(Pt 9):1965–1974. doi: 10.1099/0022-1317-71-9-1965. [DOI] [PubMed] [Google Scholar]
  23. Rittner K., Sczakiel G. Identification and analysis of antisense RNA target regions of the human immunodeficiency virus type 1. Nucleic Acids Res. 1991 Apr 11;19(7):1421–1426. doi: 10.1093/nar/19.7.1421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sarver N., Cantin E. M., Chang P. S., Zaia J. A., Ladne P. A., Stephens D. A., Rossi J. J. Ribozymes as potential anti-HIV-1 therapeutic agents. Science. 1990 Mar 9;247(4947):1222–1225. doi: 10.1126/science.2107573. [DOI] [PubMed] [Google Scholar]
  25. Schneider U., Schwenk H. U., Bornkamm G. Characterization of EBV-genome negative "null" and "T" cell lines derived from children with acute lymphoblastic leukemia and leukemic transformed non-Hodgkin lymphoma. Int J Cancer. 1977 May 15;19(5):621–626. doi: 10.1002/ijc.2910190505. [DOI] [PubMed] [Google Scholar]
  26. Schwartz S., Felber B. K., Benko D. M., Fenyö E. M., Pavlakis G. N. Cloning and functional analysis of multiply spliced mRNA species of human immunodeficiency virus type 1. J Virol. 1990 Jun;64(6):2519–2529. doi: 10.1128/jvi.64.6.2519-2529.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sczakiel G., Pawlita M. Inhibition of human immunodeficiency virus type 1 replication in human T cells stably expressing antisense RNA. J Virol. 1991 Jan;65(1):468–472. doi: 10.1128/jvi.65.1.468-472.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sczakiel G., Pawlita M., Kleinheinz A. Specific inhibition of human immunodeficiency virus type 1 replication by RNA transcribed in sense and antisense orientation from the 5'-leader/gag region. Biochem Biophys Res Commun. 1990 Jun 15;169(2):643–651. doi: 10.1016/0006-291x(90)90379-2. [DOI] [PubMed] [Google Scholar]
  29. Shibahara S., Mukai S., Morisawa H., Nakashima H., Kobayashi S., Yamamoto N. Inhibition of human immunodeficiency virus (HIV-1) replication by synthetic oligo-RNA derivatives. Nucleic Acids Res. 1989 Jan 11;17(1):239–252. doi: 10.1093/nar/17.1.239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Simons R. W. Naturally occurring antisense RNA control--a brief review. Gene. 1988 Dec 10;72(1-2):35–44. doi: 10.1016/0378-1119(88)90125-4. [DOI] [PubMed] [Google Scholar]
  31. Steuler H., Storch-Hagenlocher B., Wildemann B. Distinct populations of human immunodeficiency virus type 1 in blood and cerebrospinal fluid. AIDS Res Hum Retroviruses. 1992 Jan;8(1):53–59. doi: 10.1089/aid.1992.8.53. [DOI] [PubMed] [Google Scholar]
  32. Stockhaus J., Höfer M., Renger G., Westhoff P., Wydrzynski T., Willmitzer L. Anti-sense RNA efficiently inhibits formation of the 10 kd polypeptide of photosystem II in transgenic potato plants: analysis of the role of the 10 kd protein. EMBO J. 1990 Sep;9(9):3013–3021. doi: 10.1002/j.1460-2075.1990.tb07494.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sullenger B. A., Gallardo H. F., Ungers G. E., Gilboa E. Overexpression of TAR sequences renders cells resistant to human immunodeficiency virus replication. Cell. 1990 Nov 2;63(3):601–608. doi: 10.1016/0092-8674(90)90455-n. [DOI] [PubMed] [Google Scholar]
  34. To R. Y., Booth S. C., Neiman P. E. Inhibition of retroviral replication by anti-sense RNA. Mol Cell Biol. 1986 Dec;6(12):4758–4762. doi: 10.1128/mcb.6.12.4758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Trono D., Feinberg M. B., Baltimore D. HIV-1 Gag mutants can dominantly interfere with the replication of the wild-type virus. Cell. 1989 Oct 6;59(1):113–120. doi: 10.1016/0092-8674(89)90874-x. [DOI] [PubMed] [Google Scholar]
  36. Walder J. Antisense DNA and RNA: progress and prospects. Genes Dev. 1988 May;2(5):502–504. doi: 10.1101/gad.2.5.502. [DOI] [PubMed] [Google Scholar]
  37. Weatherall D. J. Gene therapy in perspective. Nature. 1991 Jan 24;349(6307):275–276. doi: 10.1038/349275a0. [DOI] [PubMed] [Google Scholar]
  38. Weatherall D. J. The slow road to gene therapy. Nature. 1988 Jan 7;331(6151):13–14. doi: 10.1038/331013a0. [DOI] [PubMed] [Google Scholar]
  39. Weerasinghe M., Liem S. E., Asad S., Read S. E., Joshi S. Resistance to human immunodeficiency virus type 1 (HIV-1) infection in human CD4+ lymphocyte-derived cell lines conferred by using retroviral vectors expressing an HIV-1 RNA-specific ribozyme. J Virol. 1991 Oct;65(10):5531–5534. doi: 10.1128/jvi.65.10.5531-5534.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Weintraub H. M. Antisense RNA and DNA. Sci Am. 1990 Jan;262(1):40–46. doi: 10.1038/scientificamerican0190-40. [DOI] [PubMed] [Google Scholar]
  41. Wendler I., Jentsch K. D., Schneider J., Hunsmann G. Efficient replication of HTLV-III and STLV-IIImac in human Jurkat cells. Med Microbiol Immunol. 1987;176(5):273–280. doi: 10.1007/BF00190533. [DOI] [PubMed] [Google Scholar]
  42. Wickstrom E., Simonet W. S., Medlock K., Ruiz-Robles I. Complementary oligonucleotide probe of vesicular stomatitus virus matrix protein mRNA translation. Biophys J. 1986 Jan;49(1):15–17. doi: 10.1016/S0006-3495(86)83574-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Zamecnik P. C., Goodchild J., Taguchi Y., Sarin P. S. Inhibition of replication and expression of human T-cell lymphotropic virus type III in cultured cells by exogenous synthetic oligonucleotides complementary to viral RNA. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4143–4146. doi: 10.1073/pnas.83.12.4143. [DOI] [PMC free article] [PubMed] [Google Scholar]

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