In vivo and in vitro knockdown of FREP2 gene expression in the snail Biomphalaria glabrata using RNA interference
In vivo and in vitro knockdown of FREP2 gene expression in the snail Biomphalaria glabrata using RNA interference
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DOI：
10.1016/j.dci.2005.12.004
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发表时间：
2006-01-01
影响因子：
2.9
通讯作者：
Zhang, Si-Ming
中科院分区：
文献类型：
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作者：
Jiang, Yiguo;Loker, Eric S.;Zhang, Si-Ming
本文首次报道了人类寄生虫曼氏血吸虫（Schistosoma mansoni）的中间寄主光螺（Biomphalaria glabrata）的RNA干扰。纤维蛋白原相关蛋白2 （FREP2）基因通常在暴露于遗传吸虫寄生虫（如曼氏s.m ansoni或副棘孔虫）后表达水平升高，该基因被靶向敲除。在接触吸虫前2天，将FREP2基因特定区域对应的双链RNA （dsRNA）直接注射到钉螺的血淋巴中，或加入到光裂裂芽孢杆菌胚胎细胞（Bge）和副芽孢杆菌孢子囊的共培养中。引入FREP2 dsRNA后，FREP2的表达水平显著降低，为对照组的20-30%。此外，我们能够破坏管家肌红蛋白基因的表达，进一步证实RNAi对B. glabrata的可行性。在四个FREP基因亚家族之间或FREP2与肌红蛋白之间的RNAi中未观察到交叉反应性。RNAi技术的建立为阐明被认为在宿主-寄生虫相互作用中起作用的基因的功能提供了重要的工具，特别是在光棘球绦虫与其吸虫（包括曼氏绦虫）之间。(c) 2006 Elsevier Ltd.版权所有。
RNA interference (RNAi) is reported here for the first time for Biomphalaria glabrata, the snail intermediate host for the human parasite Schistosoma mansoni. The fibrinogen-related protein 2 (FREP2) gene, normally expressed at increased levels following exposure to digenetic trematode parasites, such as S. mansoni or Echinostoma paraensei, was targeted for knockdown. Double-stranded RNA (dsRNA) corresponding to specific regions of the FREP2 gene was introduced into snails by direct injection into the hemolymph, 2 days prior to exposure to trematodes, or added to co-cultures of B. glabrata embryonic (Bge) cells and E. paraensei sporocysts. After introduction of FREP2 dsRNA, expression levels of FREP2 were significantly reduced, to 20-30% of control values. In addition, we were able to disrupt expression of the house-keeping myoglobin gene, further confirming the feasibility of RNAi for B. glabrata. Cross-reactivity in RNAi has not been observed either among four FREP gene subfamilies or between FREP2 and myoglobin. Establishment of RNAi techniques in B. glabrata provides an important tool for clarifying the function of genes believed to play a role in host-parasite interactions, specifically between B. glabrata and its trematode parasites, including S. mansoni. (c) 2006 Elsevier Ltd. All rights reserved.