Inhibition of Monkeypox virus replication by RNAi

Abstract

Orthopoxviruses cause human diseases with various severities and outcome ranging from mild conditions to death in fulminating cases. Currently, vaccination is the only protective measure against infection with these viruses and no licensed antiviral drug therapy is available. We investigated the potential of RNA interference (RNAi) in inhibiting Monkeypox virus (MPV) replication. Based on genome‐wide expression study, we selected 12 viral genes for targeting by small interference RNA (siRNA). Forty‐eight siRNA constructs were developed and evaluated in vitro for their ability to inhibit viral replication. Two genes, each targeted with four different siRNA constructs in one pool, were detrimental to viral replication. Seven siRNA constructs from these two pools, targeting either an essential gene for viral replication (A6R) or an important gene in viral entry (E8L), inhibited viral replication in cell culture by 65–95% with no apparent cytotoxicity. Further analysis using wild‐type and recombinant MPV expressing green fluorescence protein identified the most potent construct (siA6‐a) that inhibited virus replication with IC50 less than 5nM. These results identify A6R as an essential gene in MPV replication, and validate RNAi applications in developing antivirals for MPV and other Orthopoxviruses.

Document Details

Document Type
Pub Defense Publication
Publication Date
Apr 01, 2010
Source ID
10.1096/fasebj.24.1_supplement.755.5

Entities

People

  • Abdulnaser Alkhalil
  • Eric M Mucker
  • John W. Huggins
  • Peter B. Jahrling
  • Sarah Strand
  • Sofi M Ibrahim

Organizations

  • Defense Threat Reduction Agency
  • National Institute of Allergy and Infectious Diseases
  • United States Army Medical Research Institute of Infectious Diseases

Tags

Fields of Study

  • Biology

Readers

  • Infectious Disease/Epidemiology
  • Molecular Genetics

Technology Areas

  • Biotechnology
  • Biotechnology - Cancer Biotech