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A simple "soaking method" for RNA interference in the planarian Dugesia japonica

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Abstract

A simple method was developed for RNA interference (RNAi) in the planarian Dugesia japonica. The DjIFb (Dugesia japonica intermediate filament b) gene was used to evaluate the effect of RNAi because both the cDNA and an antiserum against the gene product were available. After transverse cutting at the pre- and post-pharyngeal regions, the middle part of the body fragment was soaked in water containing double-stranded RNA (dsRNA) for about 5 h and then allowed to regenerate in water. On the 5th day of regeneration, little DjIFb protein was detected in the new tissues. When the worms were cut after soaking in dsRNA water, no RNAi effect was observed, suggesting that the dsRNA was introduced through the cut surface. A high concentration of dsRNA or repeated "cutting and soaking" resulted in more effective RNAi. This simple soaking method in combination with expressed sequence tag analysis should be very useful for high-throughput analyses of gene functions in planarian regeneration.

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Fig. 1A–F.
Fig. 2A–D.

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Acknowledgements

This work was based on preliminary experiments performed by Hiroe Kurosaki, a student of the Undergraduate Life Science Research Program, Himeji Institute of Technology. We thank members of our laboratory for their support, and Dr. A. Tazaki for fruitful discussions and reading the manuscript. This work was supported in part by a Grant-in-Aid (Bio Design Program) from the Ministry of Agriculture, Forestry and Fisheries of Japan, to H.O.

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Authors and Affiliations

  1. Laboratory of Regeneration Biology, Department of Life Science, Graduate School of Science, Himeji Institute of Technology, 3-2-1 Koto, Kamigori, Akou-gun, Hyogo 678-1297, , Japan

    Hidefumi Orii, Makoto Mochii & Kenji Watanabe

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  1. Hidefumi Orii
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  2. Makoto Mochii
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  3. Kenji Watanabe
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Correspondence to Hidefumi Orii.

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Edited by N. Satoh

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Orii, H., Mochii, M. & Watanabe, K. A simple "soaking method" for RNA interference in the planarian Dugesia japonica . Dev Genes Evol 213, 138–141 (2003). https://doi.org/10.1007/s00427-003-0310-3

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  • DOI: https://doi.org/10.1007/s00427-003-0310-3

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