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Exposure to dsRNA Elicits RNA Interference in Brachionus manjavacas (Rotifera)

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Abstract

RNA interference (RNAi) is a powerful technique for functional genomics, yet no studies have reported its successful application to zooplankton. Many zooplankton, particularly microscopic metazoans of phylum Rotifera, have unique life history traits for which genetic investigation has been limited. In this paper, we report the development of RNAi methods for rotifers, with the exogenous introduction of double-stranded RNA (dsRNA) through the use of a lipofection reagent. Transfection with dsRNA for heat shock protein 90, the membrane-associated progesterone receptor, and mitogen-activated protein kinase significantly increased the proportion of non-reproductive females. Additionally, a fluorescence-based lectin binding assay confirmed the significant suppression of four of six glycosylation enzymes that were targeted with dsRNA. Suppression of mRNA transcripts was confirmed with quantitative PCR. Development of RNAi for rotifers promises to enhance the ability for assessing genetic regulation of features critical to their life history and represents a key step toward functional genomics research in zooplankton.

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Acknowledgments

This work was supported by the National Science Foundation grant BE/GenEn MCB-0412674 to TWS. We thank D. Mark Welch for providing access to the rotifer transcriptome database (GMOD) and J. Hatt and F. Loeffler for assistance with the qPCR. Undergraduates Sohee Park, Joseph Bear, Ashleigh Burns, and Michael Cray helped with PCR, dsRNA synthesis, or the bioassays.

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Authors and Affiliations

  1. School of Biology, Georgia Institute of Technology, 311 Ferst Drive, Atlanta, GA, 30332-0230, USA

    Terry W. Snell, Tonya L. Shearer & Hilary A. Smith

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  1. Terry W. Snell
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  2. Tonya L. Shearer
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  3. Hilary A. Smith
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Correspondence to Terry W. Snell.

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Snell, T.W., Shearer, T.L. & Smith, H.A. Exposure to dsRNA Elicits RNA Interference in Brachionus manjavacas (Rotifera). Mar Biotechnol 13, 264–274 (2011). https://doi.org/10.1007/s10126-010-9295-x

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  • DOI: https://doi.org/10.1007/s10126-010-9295-x

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