Production and Efficacy Test for Development of Commercial Anti-NNV RNAi Feed Additive

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Project title

Code/計畫編號

NSC102-2622-B019-002-CC2

Translated Name/計畫中文名

開發商業生產抗神經壞死病毒RNAi飼料添加劑製程及有效性測試

Project Coordinator/計畫主持人

Ming-Wei Lu

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Aquaculture

Website

https://www.grb.gov.tw/search/planDetail?id=8014046

Year

2013

Start date/計畫起

01-11-2013

Expected Completion/計畫迄

31-10-2014

Bugetid/研究經費

1000千元

ResearchField/研究領域

漁業

"神經壞死病毒(Nervous Necrosis Virus, NNV)近年來在台灣石斑魚養殖產業上一直是個不可忽視的威脅，若在石斑魚的幼苗時期發生 NNV感染往往會造成極高的死亡率。目前在研究上已有許多種傳統疫苗如死毒疫苗、次單位疫苗、及基因重組疫苗等被利用於預防 NNV的感染，但在後續的研究發現，由於石斑魚的後天免疫系統發育成熟所需時間(約孵化後 35 天)是晚於 NNV 在魚苗造成急性感染的時期，如此一來使用後天免疫的傳統策略就無法在魚苗時期達到良好的預防效果。而在本計畫欲使用的 RNA 干擾(RNA interference)技術解決這樣的問題，此技術藉由小干擾 RNA（small interfering RNA，siRNA）專一性的降解病毒性 mRNA達到抑制病毒於體內繁殖的目標。本實驗室於先前研究已將可專一性抑制NNV表現的 siRNA序列選殖到DNA載體並證實其於石斑魚細胞株或活體中抑制 NNV 感染的功效，於是在本計畫希望可以藉由和國內擁有 cGMP 認證規範及專業飼料添加劑生產技術的瑞寶基因股份有限公司合作生產高品質的 siRNA飼料添加劑試劑，投入大規模的田野試驗，除了試驗該試劑的抗 NNV感染有效性以及存放穩定性試驗，亦將完成最佳化工廠化量產 siRNA飼料添加劑試劑製程，並為未來商品化做準備。" " Nervous Necrosis Virus (NNV) is a serious disease threat in aquaculture industry recent years. NNV usually cause high mortality in infected groupers at larvae stage. There were several traditional vaccines used to against NNV infection such as inactivated vaccine, subunit vaccine and recombinant vaccine. But recent studies showed that the infection period of NNV occurred earlier than the maturation of grouper’s adaptive immunity. As this result, traditional vaccines could not provide effective protection from NNV at grouper larvae stage. However, RNA interference as new strategy in this project can provide another solution as anti-NNV strategy. By using small interfering RNA (siRNA), specific degradation of viral mRNA could suppress the replication of virus effectively. In previous study, we constructed the vector containing siRNA sequence against NNV and verified that it has effective suppression to the replication of NNV either in vitro or in vivo. This project proposes for the large scale productions of siRNA feed additive by cooperating with cGMP certified company: Reber Genetics Co., Ltd in Taiwan and proceed for large scale field tests. This cooperation can provide us to evaluate the effectiveness and stability of the siRNA additive, and further optimizes the process of siRNA feed additive production for commercialization."

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Production and Efficacy Test for Development of Commercial Anti-NNV RNAi Feed Additive

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