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. 1991 Oct;65(10):5524–5530. doi: 10.1128/jvi.65.10.5524-5530.1991

Inhibition of human immunodeficiency virus type 1 multiplication by antisense and sense RNA expression.

S Joshi 1, A Van Brunschot 1, S Asad 1, I van der Elst 1, S E Read 1, A Bernstein 1
PMCID: PMC249050  PMID: 1895401

Abstract

Human immunodeficiency virus type 1 (HIV-1) primarily infects CD4+ lymphocytes and macrophages and causes AIDS in humans. Retroviral vectors allowing neomycin phosphotransferase (npt) gene expression were engineered to express 5' sequences of HIV-1 RNA in the antisense or sense orientation and used to transform the human CD4+ lymphocyte-derived MT4 cell line. Cells expressing antisense or sense RNA to the HIV-1 tat mRNA leader sequence, as part of the 3' untranslated region of the npt mRNA, remained sensitive to HIV-1 infection. In contrast, resistance to HIV-1 infection was observed in cells expressing antisense RNA to the HIV-1 primer-binding site or to the region 5' to the primer-binding site as part of the 3' region of the npt mRNA. Cells expressing the tat mRNA leader sequence in the sense orientation as a precise replacement of the 5' untranslated region of npt mRNA were also resistant to HIV-1. These results indicate that sense and antisense approaches can be used to interfere with HIV-1 multiplication.

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Selected References

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

  1. Braddock M., Chambers A., Wilson W., Esnouf M. P., Adams S. E., Kingsman A. J., Kingsman S. M. HIV-1 TAT "activates" presynthesized RNA in the nucleus. Cell. 1989 Jul 28;58(2):269–279. doi: 10.1016/0092-8674(89)90841-6. [DOI] [PubMed] [Google Scholar]
  2. Cann A. J., Karn J. Molecular biology of HIV: new insights into the virus life-cycle. AIDS. 1989;3 (Suppl 1):S19–S34. [PubMed] [Google Scholar]
  3. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Dingwall C., Ernberg I., Gait M. J., Green S. M., Heaphy S., Karn J., Lowe A. D., Singh M., Skinner M. A., Valerio R. Human immunodeficiency virus 1 tat protein binds trans-activation-responsive region (TAR) RNA in vitro. Proc Natl Acad Sci U S A. 1989 Sep;86(18):6925–6929. doi: 10.1073/pnas.86.18.6925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ecker J. R., Davis R. W. Inhibition of gene expression in plant cells by expression of antisense RNA. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5372–5376. doi: 10.1073/pnas.83.15.5372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Edery I., Petryshyn R., Sonenberg N. Activation of double-stranded RNA-dependent kinase (dsl) by the TAR region of HIV-1 mRNA: a novel translational control mechanism. Cell. 1989 Jan 27;56(2):303–312. doi: 10.1016/0092-8674(89)90904-5. [DOI] [PubMed] [Google Scholar]
  8. Friedmann T. Progress toward human gene therapy. Science. 1989 Jun 16;244(4910):1275–1281. doi: 10.1126/science.2660259. [DOI] [PubMed] [Google Scholar]
  9. Goff S. P. Gene isolation by retroviral tagging. Methods Enzymol. 1987;152:469–481. doi: 10.1016/0076-6879(87)52055-9. [DOI] [PubMed] [Google Scholar]
  10. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  11. Izant J. G., Weintraub H. Inhibition of thymidine kinase gene expression by anti-sense RNA: a molecular approach to genetic analysis. Cell. 1984 Apr;36(4):1007–1015. doi: 10.1016/0092-8674(84)90050-3. [DOI] [PubMed] [Google Scholar]
  12. Joshi S., Van Brunschot A., Robson I., Bernstein A. Efficient replication, integration, and packaging of retroviral vectors with modified long terminal repeats containing the packaging signal. Nucleic Acids Res. 1990 Jul 25;18(14):4223–4226. doi: 10.1093/nar/18.14.4223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Joyner A. L., Bernstein A. Retrovirus transduction: generation of infectious retroviruses expressing dominant and selectable genes is associated with in vivo recombination and deletion events. Mol Cell Biol. 1983 Dec;3(12):2180–2190. doi: 10.1128/mcb.3.12.2180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  15. Lehn P. M. Gene therapy using bone marrow transplantation: a 1990 update. Bone Marrow Transplant. 1990 May;5(5):287–293. [PubMed] [Google Scholar]
  16. Levy J. A. Mysteries of HIV: challenges for therapy and prevention. Nature. 1988 Jun 9;333(6173):519–522. doi: 10.1038/333519a0. [DOI] [PubMed] [Google Scholar]
  17. Magli M. C., Dick J. E., Huszar D., Bernstein A., Phillips R. A. Modulation of gene expression in multiple hematopoietic cell lineages following retroviral vector gene transfer. Proc Natl Acad Sci U S A. 1987 Feb;84(3):789–793. doi: 10.1073/pnas.84.3.789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mann R., Mulligan R. C., Baltimore D. Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus. Cell. 1983 May;33(1):153–159. doi: 10.1016/0092-8674(83)90344-6. [DOI] [PubMed] [Google Scholar]
  19. Matsukura M., Zon G., Shinozuka K., Robert-Guroff M., Shimada T., Stein C. A., Mitsuya H., Wong-Staal F., Cohen J. S., Broder S. Regulation of viral expression of human immunodeficiency virus in vitro by an antisense phosphorothioate oligodeoxynucleotide against rev (art/trs) in chronically infected cells. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4244–4248. doi: 10.1073/pnas.86.11.4244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Melton D. A. Injected anti-sense RNAs specifically block messenger RNA translation in vivo. Proc Natl Acad Sci U S A. 1985 Jan;82(1):144–148. doi: 10.1073/pnas.82.1.144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Messing J., Crea R., Seeburg P. H. A system for shotgun DNA sequencing. Nucleic Acids Res. 1981 Jan 24;9(2):309–321. doi: 10.1093/nar/9.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Miller A. D., Buttimore C. Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production. Mol Cell Biol. 1986 Aug;6(8):2895–2902. doi: 10.1128/mcb.6.8.2895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mizuno T., Chou M. Y., Inouye M. A unique mechanism regulating gene expression: translational inhibition by a complementary RNA transcript (micRNA). Proc Natl Acad Sci U S A. 1984 Apr;81(7):1966–1970. doi: 10.1073/pnas.81.7.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Morch M. D., Joshi R. L., Denial T. M., Haenni A. L. A new 'sense' RNA approach to block viral RNA replication in vitro. Nucleic Acids Res. 1987 May 26;15(10):4123–4130. doi: 10.1093/nar/15.10.4123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Muesing M. A., Smith D. H., Capon D. J. Regulation of mRNA accumulation by a human immunodeficiency virus trans-activator protein. Cell. 1987 Feb 27;48(4):691–701. doi: 10.1016/0092-8674(87)90247-9. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Rosenberg U. B., Preiss A., Seifert E., Jäckle H., Knipple D. C. Production of phenocopies by Krüppel antisense RNA injection into Drosophila embryos. Nature. 1985 Feb 21;313(6004):703–706. doi: 10.1038/313703a0. [DOI] [PubMed] [Google Scholar]
  28. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Southgate C., Zapp M. L., Green M. R. Activation of transcription by HIV-1 Tat protein tethered to nascent RNA through another protein. Nature. 1990 Jun 14;345(6276):640–642. doi: 10.1038/345640a0. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. 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]
  32. Wigler M., Sweet R., Sim G. K., Wold B., Pellicer A., Lacy E., Maniatis T., Silverstein S., Axel R. Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell. 1979 Apr;16(4):777–785. doi: 10.1016/0092-8674(79)90093-x. [DOI] [PubMed] [Google Scholar]
  33. Zuker M., Stiegler P. Optimal computer folding of large RNA sequences using thermodynamics and auxiliary information. Nucleic Acids Res. 1981 Jan 10;9(1):133–148. doi: 10.1093/nar/9.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. van der Krol A. R., Mol J. N., Stuitje A. R. Modulation of eukaryotic gene expression by complementary RNA or DNA sequences. Biotechniques. 1988 Nov-Dec;6(10):958–976. [PubMed] [Google Scholar]
  35. von Rüden T., Gilboa E. Inhibition of human T-cell leukemia virus type I replication in primary human T cells that express antisense RNA. J Virol. 1989 Feb;63(2):677–682. doi: 10.1128/jvi.63.2.677-682.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

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