tailieunhanh - Báo cáo khoa học: In vivo RNA interference in oyster – vasa silencing inhibits germ cell development

This study investigated the potential of RNA interference, which is techni-cally challenging in bivalve mollusc species, to assess gene function in the oysterCrassostrea gigas. We designed dsRNA targeting the oyster vasa-like gene (Oyvlg), specifically expressed in oyster germ cells. In vivoinjection of oyvl-dsRNA into the gonad provokes a knockdown phenotype correspond-ing to germ cell underproliferation and prematurely arrested meiosis throu-gout the organ. | In vivo RNA interference in oyster - vasa silencing inhibits germ cell development Caroline Fabioux1 2 Charlotte Corporeau1 3 Virgile Quillien1 3 Pascal Favrel1 3 and Arnaud Huvet1 3 1 UMR 100 PE2M Ifremer-Universite de Caen Ifremer centre de Brest Plouzane France 2 UMR CNRS 6539 LEMAR Universite de Bretagne Occidentale IUEM Plouzane France 3 UMR 100 PE2M Ifremer-Universite de Caen IBFA IFR 146 ICORE Caen Cedex France Keywords Crassostrea gigas germline marine bivalve RNAi vasa Correspondence C. FabiOux UMR CNRS 6539 LEMAR Universite de Bretagne Occidentale IUEM Plouzane France Fax 33 0 2 98 49 8645 Tel 33 0 2 98 49 8744 E-mail Received 9 December 2008 revised 20 February 2009 accepted 25 February 2009 doi 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 throu-gout 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. The oyster Crassostrea gigas has stimulated a great deal of biological research as it represents a major economic resource for aquaculture production million metric tons 1 it plays a sentinel role in estuarine and coastal .

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