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<p>Nervous necrosis virus (NNV) can be transmitted vertically from parents to offspring, causing significant mortality in young fish. Therefore, effective decontamination of fertilized eggs is essential for sustainable aquaculture production. This study developed siRNA-loaded PLGA nanoparticles (NPs) for targeted silencing of NNV capsid protein (CP) and evaluated their therapeutic potential in vitro. Using a double emulsion method, siRNA-PLGA NPs were synthesized with optimized physicochemical properties: the PLGA-PEI/trichloromethane/PEG (W/O/W) system yielded monodisperse, spherical nanoparticles (~ 110&#xa0;nm post-sonication) with a high zeta potential (+ 30.91&#xa0;mV), ensuring colloidal stability and efficient cellular delivery. Overexpression of NNV CP in EPC cells induced rapid cytotoxicity, including nuclear abnormalities and cell death within 48&#xa0;h, which was fully rescued by co-administering three rationally designed CP-targeting siRNAs. Strikingly, nanoparticles resuspended in phosphate-buffered saline (PBS) exhibited a higher siRNA encapsulation efficiency, whereas those suspended in Diethyl pyrocarbonate-treated&#xa0;water showed negligible loading. These findings underscore the importance of formulation parameters in nanoparticle design and highlight the promise of RNAi-based PLGA nanotherapeutics for combating aquatic viral infections.</p>

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RNAi-based PLGA nanoparticles for targeted silencing of NNV capsid protein

  • 期刊论文

  • 发表日期：

    2025年11月1日

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  • Mingguang Mao,
  • Yaru Wang,
  • Shiyu Zhu,
  • Jielan Jiang,
  • Xiaoming Geng,
  • Xiangcheng Tang

摘要

Nervous necrosis virus (NNV) can be transmitted vertically from parents to offspring, causing significant mortality in young fish. Therefore, effective decontamination of fertilized eggs is essential for sustainable aquaculture production. This study developed siRNA-loaded PLGA nanoparticles (NPs) for targeted silencing of NNV capsid protein (CP) and evaluated their therapeutic potential in vitro. Using a double emulsion method, siRNA-PLGA NPs were synthesized with optimized physicochemical properties: the PLGA-PEI/trichloromethane/PEG (W/O/W) system yielded monodisperse, spherical nanoparticles (~ 110 nm post-sonication) with a high zeta potential (+ 30.91 mV), ensuring colloidal stability and efficient cellular delivery. Overexpression of NNV CP in EPC cells induced rapid cytotoxicity, including nuclear abnormalities and cell death within 48 h, which was fully rescued by co-administering three rationally designed CP-targeting siRNAs. Strikingly, nanoparticles resuspended in phosphate-buffered saline (PBS) exhibited a higher siRNA encapsulation efficiency, whereas those suspended in Diethyl pyrocarbonate-treated water showed negligible loading. These findings underscore the importance of formulation parameters in nanoparticle design and highlight the promise of RNAi-based PLGA nanotherapeutics for combating aquatic viral infections.