Enhanced expression and HIV- 1 inhition of chimeric tRNALys3-Ribozymes under dual U6 snRNA and tRNA promoters

Zongli Chang, Shawn Westaway, Shirley Li, John A. Zaia, John J. Rossi, Lisa J. Scherer

Research output: Contribution to journalArticlepeer-review

15 Scopus citations


We previously demonstrated that chimeric tRNALys3-ribozymes targeting the primer binding site of HIV produced virions with reduced infectivity. To further enhance the anti-HIV efficiency of these ribozymes by increasing their level of transcription, we designed several tRNALys3 promoter variants and compared their expression levels from the internal tRNALys3 promoters and also from an exogenous human U6 snRNA promoter. The dual U6/tRNA promoter constructs gave rise to much higher levels of expression than constructs that used only an internal tRNA promoter. The most abundant expression is produced when a U6 promoter drives a chimeric tRNALys3-ribozyme containing a mutation in the tRNA B box. As detected by fluorescent in situ hybridization, transcripts from a construct with the tRNA promoter alone localized strictly to the cytoplasm, whereas transcripts from dual U6/tRNA promoter were present in both the cytoplasm and the nucleus. Inhibition of HIV-1 correlates well with expression levels of the chimeric constructs. The results presented demonstrate that U6 and tRNA promoters can be placed in tandem for high-level expression of small RNA therapeutic transcripts.

Original languageEnglish (US)
Pages (from-to)481-489
Number of pages9
JournalMolecular Therapy
Issue number4
StatePublished - Oct 1 2002


  • Gene therapy
  • Human immunodeficiency virus
  • Primer binding site
  • Promoter
  • RNA polymerase III
  • Ribozyme
  • Subcellular localization
  • U6 small nuclear RNA (U6 snRNA)
  • tRNA
  • tRNA

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Pharmacology
  • Drug Discovery


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