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Plants rely on a multi-layered innate immune system to defend against abiotic and biotic, with RNA interference (RNAi) serving as a core antiviral mechanism. RNAi generates small interfering RNAs (siRNAs) that direct sequence-specific degradation and translational inhibition of viral RNAs. However, conventional strategies involving insertion of viral genes into the plant genome pose potential biosafety concerns. By contrast, RNAi-mediated silencing of endogenous host susceptibility genes offers distinct advantages, especially when combined with natural recessive resistance alleles. This chapter describes a detailed protocol for hairpin RNA (hpRNA) construct design and application, providing a practical reference for researchers screening and identifying host factors required for plant virus infection.

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Silencing Host Factors with Hairpin RNA for Virus Resistance

  • 实验方案

  • 发表日期：

    2026年

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  • Penghuan Rui,
  • Jianping Chen,
  • Fei Yan,
  • Guanwei Wu

摘要

Plants rely on a multi-layered innate immune system to defend against abiotic and biotic, with RNA interference (RNAi) serving as a core antiviral mechanism. RNAi generates small interfering RNAs (siRNAs) that direct sequence-specific degradation and translational inhibition of viral RNAs. However, conventional strategies involving insertion of viral genes into the plant genome pose potential biosafety concerns. By contrast, RNAi-mediated silencing of endogenous host susceptibility genes offers distinct advantages, especially when combined with natural recessive resistance alleles. This chapter describes a detailed protocol for hairpin RNA (hpRNA) construct design and application, providing a practical reference for researchers screening and identifying host factors required for plant virus infection.