RNA interference in marine and freshwater sponges: actin knockdown in Tethya wilhelma and Ephydatia muelleriby ingested dsRNA expressing bacteria

BMC Biotechnology

Methodology Article

Ian P Winters¹  April L Hill¹  Malcolm S Hill¹  Dora Posfai¹  Ajna S Rivera²  Elizabeth S Danka³  Brandon Cieniewicz⁴  Scott W Knight⁵  Gert Wörheide⁶  Jörg U Hammel⁷  Michael Nickel⁷  Dennis V Lavrov⁸  Karri M Haen⁸ 

[1] Department of Biology, University of Richmond, Richmond, VA, USA;Department of Biology, University of Richmond, Richmond, VA, USA;Biology Department, College of the Pacific, Stockton, CA, USA;Department of Biology, University of Richmond, Richmond, VA, USA;Molecular Cell Biology, Washington University, St. Louis, MO, USA;Department of Biology, University of Richmond, Richmond, VA, USA;Molecular and Cellular Biology, Stony Brook University, Stony Brook, NY, USA;Department of Biology, University of Richmond, Richmond, VA, USA;Sigma-Aldrich Biotechnology, St. Louis, MO, USA;Department of Earth and Environmental Sciences & GeoBio-CenterLMU, Ludwig-Maximilians-University Munich, Germany;Institut für Spezielle Zoologie und Evolutionsbiologie mit Phyletischem Museum, Friedrich-Schiller-Universität, Jena, Germany;Iowa State University, Ames, IA, USA;

关键词: Sponge;    Sponge Tissue;    Freshwater Sponge;    Phalloidin Staining;    Cytoplasmic Actin;   

DOI  :  10.1186/1472-6750-11-67

 received in 2010-12-09, accepted in 2011-06-16,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundThe marine sponge Tethya wilhelma and the freshwater sponge Ephydatia muelleri are emerging model organisms to study evolution, gene regulation, development, and physiology in non-bilaterian animal systems. Thus far, functional methods (i.e., loss or gain of function) for these organisms have not been available.ResultsWe show that soaking developing freshwater sponges in double-stranded RNA and/or feeding marine and freshwater sponges bacteria expressing double-stranded RNA can lead to RNA interference and reduction of targeted transcript levels. These methods, first utilized in C. elegans, have been adapted for the development and feeding style of easily cultured marine and freshwater poriferans. We demonstrate phenotypic changes result from 'knocking down' expression of the actin gene.ConclusionThis technique provides an easy, efficient loss-of-function manipulation for developmental and gene regulatory studies in these important non-bilaterian animals.

【 授权许可】

Unknown   
© Rivera et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.