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Artificial MicroRNA-Mediated Inhibition of Japanese Encephalitis Virus Replication in Neuronal Cells

Himani Sharma1,2, Aarti Tripathi1, Bharti Kumari1

  • 11 Vaccine and Infectious Disease Research Center (VIDRC), Translational Health Science and Technology Institute (THSTI), Faridabad, India.

Nucleic Acid Therapeutics
|November 21, 2018
PubMed

Insights

Artificial microRNAs effectively inhibit Japanese encephalitis virus (JEV) replication by targeting conserved 3' untranslated regions. This RNA interference approach significantly reduces viral RNA, proteins, and infectious particles, offering a promising antiviral strategy.

Area of Science:

  • Virology
  • Molecular Biology
  • Antiviral Therapy

Background:

  • Artificial microRNAs (amiRNAs) are emerging as a potent strategy for inhibiting viral replication.
  • Japanese encephalitis virus (JEV) poses a significant public health threat, necessitating novel therapeutic approaches.

Purpose of the Study:

  • To evaluate the efficacy of amiRNA-mediated inhibition against Japanese encephalitis virus (JEV).
  • To assess the potential of targeting the 3' untranslated region (3'UTR) of JEV for antiviral therapy.

Main Methods:

  • Designed three amiRNA sequences targeting the conserved 3'UTR of JEV.
  • Tested amiRNA binding affinity in vitro using a luciferase reporter assay.
  • Assessed amiRNA efficacy in neuronal cells through viral RNA quantification, protein analysis (NS1), and plaque reduction assays.

Main Results:

  • Transient amiRNA transfection was non-toxic and minimally induced interferon-stimulated genes.
  • Two amiRNAs (amiRNA #1 and #2) significantly reduced intracellular viral RNA and NS1 protein levels.
  • Infectious viral particle release was diminished by up to 95% in the supernatant.

Conclusions:

  • RNA interference utilizing amiRNAs targeting conserved JEV 3'UTR regions is an effective antiviral strategy.
  • This approach shows promise for developing improved nucleic acid-based inhibitors against JEV.
Keywords:
3′UTRJEVartificial microRNAreplication

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