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Updated: Apr 30, 2026

Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction
Published on: November 1, 2018
Interferon-mediated ISG15 conjugation restricts dengue virus 2 replication.
Takayuki Hishiki1, Qi'En Han2, Kei-ichiro Arimoto3
1Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, 14 Medical Drive,Singapore 117599, Singapore; Laboratory of Primate Model, Experimental Research Center for Infectious Diseases, Institute for Virus Research, Kyoto University, Kyoto 606-8517, Japan.
Interferon-inducible ISGylation, a modification by ISG15, suppresses Dengue virus 2 (DENV-2) release. ISG15 acts as a host antiviral factor, reducing infectious DENV-2 particle production.
Area of Science:
- Immunology
- Virology
- Molecular Biology
Background:
- ISGylation is an interferon-stimulated, ubiquitin-like modification mediated by ISG15.
- ISGylation plays a role in the host's antiviral defense mechanisms.
Purpose of the Study:
- To investigate the functional role of ISGylation in Dengue virus 2 (DENV-2) replication.
- To determine if ISG15 acts as a host antiviral factor against DENV-2.
Main Methods:
- Overexpression of ISG15 and a conjugation-defective ISG15 mutant.
- RNA interference to deplete endogenous ISG15.
- Analysis of extracellular and intracellular viral RNA and infectious virus release.
- Identification of DENV-2 proteins targeted by ISGylation.
Main Results:
- ISG15 overexpression significantly suppressed extracellular infectious DENV-2 release and decreased virus infectivity.
- Intracellular viral RNA levels remained unaffected by ISG15 overexpression.
- ISG15 depletion partially restored DENV-2 RNA levels in interferon-treated cells.
- DENV-2 NS3 and NS5 proteins were identified as targets of ISGylation.
Conclusions:
- Interferon-inducible ISGylation inhibits DENV-2 particle release.
- ISG15 is a key mediator of the interferon-induced antiviral response against DENV-2.
- ISG15 functions as a host antiviral factor against DENV-2 infection.
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