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浙大解析病毒miRNA在體內(nèi)調(diào)控宿主抗病毒細(xì)胞凋亡分子機(jī)制
點擊次數(shù)：1079 發(fā)布時間：2014-2-11
病毒，尤其是DNA病毒，會產(chǎn)生自身的microrna(miRNA)來調(diào)控宿主和病毒基因的表達(dá)。由于其在病毒—宿主相互作用中的重要作用，近年來病毒miRNA已引起了廣泛的關(guān)注與研究，然而人們對無脊椎動物病毒miRNA的了解卻十分有限。至今，大多數(shù)對病毒miRNA的研究是在細(xì)胞系中進(jìn)行的，但是病毒miRNA在細(xì)胞系中發(fā)揮的作用可能與其在宿主體內(nèi)的作用差異巨大。
在本項研究中，浙江大學(xué)生命科學(xué)學(xué)院章曉波教授課題組對對蝦體內(nèi)的白斑綜合癥病毒(WSSV)編碼的miRNA進(jìn)行了表征分析，揭示了病毒miRNA調(diào)控宿主抗病毒的機(jī)制*，相關(guān)研究成果發(fā)表在12月刊的Journal of Virology。
研究人員采用高通量測序技術(shù)對WSSV侵染的對蝦(Marsupenaeus japonicus)小RNA組進(jìn)行分析(小RNA測序由聯(lián)川生物提供技術(shù)服務(wù))，并結(jié)合先前的研究數(shù)據(jù)，共鑒定出89個假定的WSSV miRNA。
利用μParaflo®微流體定制miRNA芯片分析(定制miRNA芯片合成檢測由聯(lián)川生物提供技術(shù)服務(wù))和Northern雜交驗證揭示出病毒miRNA的表達(dá)在對蝦體內(nèi)具有組織特異性。進(jìn)一步實驗表明，病毒miRNA WSSV-miR-N24可以靶向調(diào)控對蝦的caspase 8基因，從而抑制對蝦體內(nèi)血細(xì)胞的凋亡。此項研究報道了病毒miRNA在體內(nèi)調(diào)控宿主抗病毒細(xì)胞凋亡的分子機(jī)制。
原文摘要：
Involvement of viral miRNA in the regulation of antiviral apoptosis in shrimp
Tianzhi Huang, Yalei Cui and Xiaobo Zhang
Viruses, in particular DNA viruses, generate their microRNAs (miRNAs) to control expressions of host and viral genes. Due to their essential roles in virus-host interactions, viral miRNAs have attracted extensive investigations in recent years. Up to date, however, most studies on viral miRNAs are conducted in cell lines. In this study, the viral miRNAs from white spot syndrome virus (WSSV) were characterized in shrimp in vivo. On the basis of our previous study and small RNA sequencing in this study, a total of 89 putative WSSV miRNAs were identified. As revealed by miRNA microarray analysis and Northern blots, the expressions of viral miRNAs were tissue-specific in vivo. The results indicated that the viral miRNA WSSV-miR-N24 could target the shrimp caspase 8 gene, and further repressed the apoptosis of shrimp hemocytes in vivo. As a result, the WSSV copies in shrimp in vivo were significantly increased compared with the control (WSSV only). Therefore, our study presented the first report on the in vivomolecular events of viral miRNA in the antiviral apoptosis.