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Viral envelope protein 53R gene highly specific silencing and iridovirus resistance in fish Cells by AmiRNA

Yu-Sin Kim1, Fei Ke, Xiao-Ying Lei

  • 1State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China.

Plos One
|April 30, 2010
PubMed
Abstract

Insights

This study introduces artificial microRNAs (amiRNAs) to silence the frog virus Rana grylio virus (RGV) envelope protein 53R in fish cells. This novel approach effectively inhibits RGV infection and virion assembly, offering a new strategy for antiviral resistance.

Area of Science:

  • Virology
  • Molecular Biology
  • Biotechnology

Background:

  • The frog virus Rana grylio virus (RGV) is an aquatic animal pathogen.
  • The RGV envelope protein 53R is crucial for virus assembly.
  • RNA interference (RNAi) has not been previously reported for inhibiting aquatic animal virus envelope proteins.

Purpose of the Study:

  • To develop artificial microRNAs (amiRNAs) targeting the RGV 53R envelope protein gene.
  • To investigate the efficacy of amiRNAs in inhibiting RGV replication and virion assembly in fish cells.
  • To establish a novel RNAi-based antiviral strategy for aquatic animals.

Main Methods:

  • Artificial microRNAs (amiRNAs) targeting RGV 53R were designed and incorporated into pre-miRNA155 vectors.
  • These vectors were transfected into fish cell lines to express amiRNAs (amiR-53Rs).
  • RGV 53R expression, viral load, and virion assembly were analyzed using real-time PCR and electron microscopy.

Main Results:

  • The amiRNA amiR-53R-1 effectively silenced the RGV 53R gene expression.
  • Viral infection was suppressed by 58% with amiR-53R-1 targeting a specific region of RGV 53R.
  • Electron microscopy confirmed defects or reduced capacity in virion assembly correlated with amiR-53R-1 expression.

Conclusions:

  • This study presents the first report of an amiRNA targeting an iridovirus envelope protein gene.
  • amiR-53R-1 demonstrates significant potential for inhibiting RGV infection and virion assembly in fish.
  • This research establishes a foundation for developing RNAi-based antiviral therapies for aquatic viral diseases.

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