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Inhibition of Viral Multiplication in Acute and Chronic Stages of Infection by Ribozymes Targeted against the Polymerase Gene of Mouse Hepatitis Virus

  • Akihiko Maeda
  • Tetsuya Mizutani
  • Masanobu Hayashi
  • Kozue Ishida
  • Tomomasa Watanabe
  • Shigeo Namioka
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 380)

Abstract

Two hammerhead ribozymes targeted against the polymerase gene of mouse hepatitis virus (MHV), which consisted of 22-nucleotide (nt) ribozyme core sequences and antisense sequences of different lengths, 243-nt (S-ribozyme) and 926-nt (L-ribozyme), were tested for their inhibitory effects on viral multiplication. Vectors that expressed the ribozymes were transfected into mouse DBT cells and several resulting cell lines constitutively expressing the ribozymes were selected and examined for intracellular MHV multiplication in acute and chronic stages of infection. The production of infectious progeny viral particles was significantly reduced in the transfected cell lines expressing either the S-ribozyme or L-ribozyme in acute infection. Although the in vitro cleavage process of the L-ribozyme was slower than that of the S-ribozyme, no difference was observed in inhibitory effects on MHV multiplication between S-and L-ribozymes in the transfected cells. In the transfected cells expressing L-ribozymes, production of viral particles was also inhibited in the chronic stage of MHV infection.

Keywords

Chronic Stage Cleavage Reaction Polymerase Gene Hammerhead Ribozyme Mouse Hepatitis Virus 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Cech, T.R. The chemistry ofself-splicing RNA and RNA enzymes. Science 1987; 236: 1532–1539.PubMedCrossRefGoogle Scholar
  2. 2.
    Haseloff,J., Gerlach, W.L.Nature (Lond.) Simple RNA enzymes with new and highly specific endonuclease activity. 1988; 334: 585–591.Google Scholar
  3. 3.
    Uhlenbeck, O.C. A small catalyticoligoribonucleotide. Nature 1987; 328: 596–600.PubMedCrossRefGoogle Scholar
  4. 4.
    Sarver, N.,Cantin, E.M., Chang, P.S., Zaia, J.A., Stephens, D.A., Rossi, J.J. Ribozymes aspotencial anti-HIV-1 therapeutic agents.Science 1989; 247: 1222–1225.CrossRefGoogle Scholar
  5. 5.
    Xing, Z.,Whitton, J.L. An anti-lymphocytic choriomeningitis virus ribozyme expressed intissue culture cellsdiminishes viral RNA levels and leads to a reduction in infectious virus yield.J. Virol. 1993; 67: 1840–1847.PubMedGoogle Scholar
  6. 6.
    Denman, R.B.,Purow, B., Rubenstein, R., Miller, D.L. Hammerhead ribozyme cleavage of hamsterprion pre-mRNA incomplex cell-free model systems. Biochem. Biophys. Res. Commun. 1992; 186:1171–1177.PubMedCrossRefGoogle Scholar
  7. 7.
    Hirano, N.,Fujiwara, K., Hino, S., Matumoto, M. Replication and plaque formation of mousehepatitis virus (MHV-2) in mouse cell line DBT culture. Arch. Ges. Virusforsch. 1974; 44: 298–302.PubMedCrossRefGoogle Scholar
  8. 8.
    Makino, S.,Stohlman, S.A., Lai, M.M.C. Leader sequences of murine coronavirus mRNAs can befreely reassorted: Evidence for the role of free leader RNA in transcription.Proc. Natl. Acad. U.S.A. 1986; 83: 4204–4208.CrossRefGoogle Scholar
  9. 9.
    Hirano, N.,Goto, N., Makino, S., Fujiwara, K. Persistent infection with mouse hepatitisvirus JHM strain in DBT cell culture. Adv.Exp. Med. Biol. 1981; 142: 301–308.PubMedGoogle Scholar
  10. 10.
    Kim, D.W.,Uetsuki, T., Kajiro, Y., Yamaguchi, N., Sugano, S. Use of the human elongationfacter la promoteras a versatile and efficient expression system. Gene 1990; 91: 217–223.PubMedCrossRefGoogle Scholar
  11. 11.
    Graham, F.L.,Van der Eb, A.J. A new technique for theassay of infectivity of human adenovirus 5 DNA. Virology 1973; 52: 456–467.PubMedCrossRefGoogle Scholar
  12. 12.
    Asanaka, M.,Lai, M.M.C. Cell fusion studies identified multiple factors involved in mousehepatitis virus entry. Virology 1993; 197:732–741.PubMedCrossRefGoogle Scholar
  13. 13.
    Mizzen, L.,Cheley, S., Rao, M., Wolf, R., Anderson, R. Fusion resistance and decreasedinfectability as majorhost cell determinants of coronavirus persistence. Virology 1983; 128:407–417.PubMedCrossRefGoogle Scholar
  14. 14.
    . Brayton, P.R., Lai, M.M.C.,Patton, C.D., Stohlman, S.A. Characterization of two RNA polymerase activities induced by mouse hepatitis virus. J.Virol.1982; 42: 847–853.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • Akihiko Maeda
    • 1
  • Tetsuya Mizutani
    • 1
  • Masanobu Hayashi
    • 1
  • Kozue Ishida
    • 1
  • Tomomasa Watanabe
    • 1
  • Shigeo Namioka
    • 1
  1. 1.Department of Laboratory Animal Science, Faculty of Veterinary MedicineHokkaido UniversitySapporoJapan

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