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RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
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Related Experiment Video

Updated: May 30, 2025

Monitoring Activation of the Antiviral Pattern Recognition Receptors RIG-I And PKR By Limited Protease Digestion and Native PAGE
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Dengue Virus Replicative-Form dsRNA Is Recognized by Both RIG-I and MDA5 to Activate Innate Immunity.

Sichao Ye1,2, Yisha Liang1,2,3, Yu Chang1,3

  • 1CAS Key Laboratory of Molecular Virology and Immunology, Shanghai Institute of Immunity and Infection, Chinese Academy of Sciences, Shanghai, China.

Journal of Medical Virology
|January 28, 2025
PubMed
Summary

Dengue virus (DENV) infection triggers an interferon response mediated by RIG-I like receptors (RLRs), primarily RIG-I and MDA5. Researchers identified DENV

Keywords:
MDA5RIG‐Idengue viruspathogen‐associated molecular patternsreplicative form

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Area of Science:

  • Immunology
  • Virology
  • Molecular Biology

Background:

  • RIG-I like receptors (RLRs) are crucial cytosolic sensors for RNA virus infections, initiating innate immune responses.
  • The specific pathogen-associated molecular patterns (PAMPs) of Dengue virus (DENV) recognized by RLRs remain poorly understood.
  • Distinct RLRs, RIG-I and MDA5, are generally thought to recognize different types of viral RNA structures.

Purpose of the Study:

  • To elucidate the molecular basis of DENV PAMP recognition by host RLRs.
  • To investigate the roles of RIG-I and MDA5 in the DENV-induced interferon response.
  • To characterize the specific DENV RNA structures that activate RLRs.

Main Methods:

  • Assessed DENV infection-induced interferon response in the presence of RIG-I and MDA5.
  • Purified DENV PAMP RNA from infected cells.
  • Confirmed DENV PAMP identity by in vitro reconstitution of viral replicative-form RNA.

Main Results:

  • DENV infection activates interferon production dependent on both RIG-I and MDA5, with RIG-I being predominant.
  • The DENV PAMP was identified as full-length double-stranded RNA with 5' triphosphate (5'ppp) modifications.
  • This identified DENV PAMP likely represents the viral replicative-form RNA.

Conclusions:

  • This study defines the molecular interaction between DENV PAMPs and RLRs, clarifying innate immune recognition.
  • The findings reveal that the same PAMP moiety can be recognized by different RLRs, highlighting a nuanced innate immunity mechanism.
  • The research contributes to understanding host defense against RNA viruses like DENV.