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Systemic RNA Interference in Planarians by Feeding of dsRNA Containing Bacteria

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Planarian Regeneration

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1774))

Abstract

RNA interference (RNAi) is currently the only method available in planaria for assessing the function of particular genes. We describe here a method for performing body-wide gene knockdown, relying on dsRNA production in bacteria and subsequent delivery to planaria by feeding a liver–bacteria mixture. This method is ideal for screening many genes in parallel, in a cost-effective and reliable manner. We also describe a ligation-independent cloning strategy, which is used to rapidly transfer single genes into an RNAi vector that is also appropriate for downstream applications such as in situ hybridizations. Together, these protocols represent useful components of the current planarian molecular tool kit.

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Author information

Authors and Affiliations

  1. Cornell University, Ithaca, NY, USA

    Carolyn E. Adler

  2. Howard Hughes Medical Institute, Stowers Institute for Medical Research, Kansas, MO, USA

    Alejandro Sánchez Alvarado

Authors
  1. Carolyn E. Adler
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  2. Alejandro Sánchez Alvarado
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Corresponding author

Correspondence to Alejandro Sánchez Alvarado .

Editor information

Editors and Affiliations

  1. Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany

    Jochen C. Rink

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Adler, C.E., Alvarado, A.S. (2018). Systemic RNA Interference in Planarians by Feeding of dsRNA Containing Bacteria. In: Rink, J. (eds) Planarian Regeneration. Methods in Molecular Biology, vol 1774. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7802-1_17

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  • DOI: https://doi.org/10.1007/978-1-4939-7802-1_17

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7800-7

  • Online ISBN: 978-1-4939-7802-1

  • eBook Packages: Springer Protocols

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