In vivo RNA interference in oyster -vasa silencing inhibits germ cell development
In vivo RNA interference in oyster -vasa silencing inhibits germ cell development
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DOI：
10.1111/j.1742-4658.2009.06982.x
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发表时间：
2009-05-01
期刊：
影响因子：
5.4
通讯作者：
Huvet, Arnaud
中科院分区：
文献类型：
--
作者：
Fabioux, Caroline;Corporeau, Charlotte;Huvet, Arnaud
本研究探讨了RNA干扰的潜力，这是技术上具有挑战性的双壳类软体动物物种，以评估基因功能的牡蛎长牡蛎。我们设计了针对牡蛎vasa样基因（Oyvlg）的dsRNA，该基因在牡蛎生殖细胞中特异性表达。在体内将oyvl-dsRNA注射到性腺中引起对应于生殖细胞增殖不足的敲低表型，并且过早地阻止了器官的减数分裂。观察到的最严重的表型是不育。这种敲低表型与Oyvlg mRNA水平降低39%至87%以及OYVLG蛋白质强烈降低至不可检测的水平相关。因此，Oyvlg似乎是必不可少的生殖细胞的发育，特别是有丝分裂增殖和早期减数分裂。我们的研究结果首次表明，在体内RNA干扰有效地在一个双壳类物种，打开功能遗传学研究的主要前景。
This study investigated the potential of RNA interference, which is technically challenging in bivalve mollusc species, to assess gene function in the oyster Crassostrea gigas. We designed dsRNA targeting the oyster vasa-like gene (Oyvlg), specifically expressed in oyster germ cells. In vivo injection of oyvl-dsRNA into the gonad provokes a knockdown phenotype corresponding to germ cell underproliferation and prematurely arrested meiosis througout the organ. The most severe phenotype observed is sterile. This knockdown phenotype is associated with a decrease in Oyvlg mRNA level of between 39% and 87%, and a strong reduction in OYVLG protein, to an undetectable level. Therefore, Oyvlg appears to be essential for germ cell development in Crassostrea gigas, particularly for mitotic proliferation and early meiosis. Our results demonstrate for the first time that in vivo RNA interference works efficiently in a bivalve species, opening major perspectives for functional genetic studies.