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Use of a nonviral vector to express a chimeric trna-ribozyme against lymphocytic choriomeningitis virus: Cytoplasmic accumulation of a catalytically competent transcript but minimal antiviral effect

Academic Article
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Overview

authors

  • Gebhard, J. R.
  • Perry, C. M.
  • Mahadeviah, S.
  • Whitton, J. Lindsay

publication date

  • 1997

journal

  • Antisense & Nucleic Acid Drug Development  Journal

abstract

  • RNA polymerase III promoters direct the ubiquitous, high-level, expression of small, stable RNAs such as tRNAs, and thus are attractive candidates for achieving stable expression of small therapeutic (e.g., antiviral) molecules, such as ribozymes or antisense RNAs. In this article, we describe the use of a nonviral vector containing a tRNA promoter to express an antilymphocytic choriomeningitis virus (LCMV) ribozyme (tRNA-Rib5). The chimeric tRNA-ribozyme is specifically and efficiently transcribed by pol III in cell-free extracts, and the resulting transcript has appropriate ribozyme activity. In tissue culture studies, high levels of chimeric transcripts were readily detectable and were transported to the cytoplasm, the site of LCMV replication. Despite accumulation of tRNA-Rib5 in the cytoplasm of stably transformed cell clones, antiviral effects were minimal or absent. The implications of these findings and the potential use of this vector system for in vivo studies requiring the delivery of small molecules are discussed.

subject areas

  • 3T3 Cells
  • Animals
  • Antiviral Agents
  • Cercopithecus aethiops
  • Chimera
  • Clone Cells
  • Cytoplasm
  • Gene Expression
  • Genetic Vectors
  • Lymphocytic choriomeningitis virus
  • Mice
  • Mice, Inbred BALB C
  • Oligonucleotides, Antisense
  • Plasmids
  • Promoter Regions, Genetic
  • RNA Polymerase III
  • RNA, Catalytic
  • RNA, Transfer, Met
  • Transcription, Genetic
  • Vero Cells
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Identity

International Standard Serial Number (ISSN)

  • 1087-2906

Digital Object Identifier (DOI)

  • 10.1089/oli.1.1997.7.3

PubMed ID

  • 9055033
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Additional Document Info

start page

  • 3

end page

  • 11

volume

  • 7

issue

  • 1

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