Targeted cleavage of hepatitis E virus 3′ end RNA mediated by hammerhead ribozymes inhibits viral RNA replication

Bandi Sriram, Deepshi Thakral, Subrat Kumar Panda

15 Scopus citations


The 3′ end of hepatitis E virus (HEV) contains cis-acting regulatory element, which plays an important role in viral replication. To develop specific replication inhibitor at the molecular level, mono- and di-hammerhead ribozymes (Rz) were designed and synthesized against the conserved 3′ end sequences of HEV, which cleave at nucleotide positions 7125 and 7112/7125, respectively. Di-hammerhead ribozyme with two catalytic motifs in tandem was designed to cleave simultaneously at two sites spaced 13 nucleotides apart, which increases the overall cleavage efficiency and prevents the development of escape mutants. Specific cleavage products were obtained with both the ribozymes in vitro at physiological conditions. The inactive control ribozymes showed no cleavage. The ribozymes showed specific inhibition of HEV 3′ end fused-luciferase reporter gene expression by ∼37 and ∼60%, respectively in HepG2 cells. These results demonstrate a feasible approach to inhibit the HEV replication to a limited extent by targeting the cis-acting 3′ end of HEV with hammerhead ribozymes.

Original languageEnglish (US)
Pages (from-to)350-358
Number of pages9
Issue number2
Publication statusPublished - Aug 1 2003
Externally publishedYes



  • Cis-acting
  • Gene expression
  • Gene therapy
  • Trans-acting
  • Transient expression

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

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