Characterization of Argonaute cDNA from Penaeus monodon and implication of its role in RNA interference
Characterization of Argonaute cDNA from Penaeus monodon and implication of its role in RNA interference
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DOI：
10.1016/j.bbrc.2008.01.031
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发表时间：
2008-03-21
影响因子：
3.1
通讯作者：
Panyim, Sakol
中科院分区：
文献类型：
--
作者：
Dechklar, Manasave;Udomkit, Apinunt;Panyim, Sakol
RNA干扰技术是近年来发展起来的一种治疗病毒性疾病的有效方法。然而，在斑节对虾中发挥RNAi作用的蛋白质组分尚未被鉴定。在这里，我们报告的cDNA编码Argonaute，在斑节对虾RNAi途径的主要成分的克隆和功能特性。斑节对虾Argonaute（Pem-AGO）具有PAZ和PIWI两个特征结构域。在对虾原代培养的Oka细胞中，针对PAZ编码序列的双链RNA（dsRNA）可以抑制Pem-ago的表达。Pem-ago耗尽的细胞显示出受损的RNAi，因为内源基因的表达从这些细胞中dsRNA介导的沉默中被拯救。我们的研究结果表明，Pem-ago是斑节对虾有效RNAi所必需的，从而确定了对虾RNAi机制的第一个蛋白质组分。(c)2008年爱思唯尔公司All rights reserved.
RNA interference (RNAi) has recently become a promising strategy for therapeutic of several viral diseases including those in the black tiger shrimp Penaeus monodon. However, the protein components that play role in RNAi in P. monodon have not yet been identified. Here, we report the cloning and functional characterization of a cDNA encoding Argonaute, a principal constituent of RNAi pathway in P. monodon. P. monodon's Argonaute (Pem-AGO) exhibited the two signature domains, PAZ and PIWI. Substantial level of Pem-ago expression could be suppressed by double-stranded RNA (dsRNA) that targeted PAZ coding sequence in shrimp primary culture of Oka cells. The Pem-ago depleted cells showed impaired RNAi as the expression of an endogenous gene was rescued from the dsRNA-mediated silencing in these cells. Our results imply that Pem-ago is required for effective RNAi in P. monodon and thus identify the first protein constituent of RNAi machinery in penaeid shrimp. (c) 2008 Elsevier Inc. All rights reserved.