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Preparation and Use of a Yeast shRNA Delivery System for Gene Silencing in Mosquito Larvae

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Insect Genomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1858))

Abstract

The mosquito genome projects facilitated research in new facets of mosquito biology, including functional genetic studies in the dengue and Zika virus vector Aedes aegypti and the primary African malaria vector Anopheles gambiae. RNA interference (RNAi) has facilitated gene silencing experiments in both of these disease vector mosquito species and could one day be applied as a new method of vector control. Here, we describe a procedure for the genetic engineering of Saccharomyces cerevisiae (baker’s yeast) that express short hairpin RNA (shRNA) corresponding to mosquito target genes of interest. Following cultivation, which facilitates inexpensive propagation of shRNA, the yeast is inactivated and prepared in a ready-to-use dry tablet formulation that is fed to mosquito larvae. Ingestion of the yeast tablets results in effective larval target gene silencing. This technically straightforward and affordable technique may be applicable to a wide variety of mosquito species and potentially to other arthropods that feed on yeast.

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Acknowledgments

We thank Ping Li, Longhua Sun, Chien-Wei Chen, Yingying Chen, Joanne Cunningham, and Diane Lovin for their technical assistance during development of these methodologies. Development of this protocol was funded by a United States Agency for International Development Award AID-OAA-F-16-00097 to MDS and by NIH/NIAID Award 1 R21 AI128116-01 to MDS, DWS, and NW.

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Author notes

  1. Keshava Mysore and Limb K. Hapairai contributed equally to this work.

Authors and Affiliations

  1. Department of Medical and Molecular Genetics, Indiana University School of Medicine, South Bend, IN, USA

    Keshava Mysore, Limb K. Hapairai, Jacob S. Realey, David W. Severson & Molly Duman-Scheel

  2. Eck Institute for Global Health, University of Notre Dame, Notre Dame, IN, USA

    Keshava Mysore, Limb K. Hapairai, Na Wei, Jacob S. Realey, Nicholas D. Scheel, David W. Severson & Molly Duman-Scheel

  3. Department of Civil and Environmental Engineering and Earth Sciences, University of Notre Dame, Notre Dame, IN, USA

    Na Wei

  4. Department of Biological Sciences, University of Notre Dame, Notre Dame, IN, USA

    Nicholas D. Scheel, David W. Severson & Molly Duman-Scheel

Authors
  1. Keshava Mysore
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  2. Limb K. Hapairai
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  3. Na Wei
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  4. Jacob S. Realey
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  5. Nicholas D. Scheel
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  6. David W. Severson
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  7. Molly Duman-Scheel
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Corresponding author

Correspondence to Molly Duman-Scheel .

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Editors and Affiliations

  1. Division of Biology, Kansas State University, Manhattan, NY, USA

    Susan J. Brown

  2. Department of Biological Sciences, University of Notre Dame, Notre Dame, IN, USA

    Michael E. Pfrender

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Mysore, K. et al. (2019). Preparation and Use of a Yeast shRNA Delivery System for Gene Silencing in Mosquito Larvae. In: Brown, S., Pfrender, M. (eds) Insect Genomics. Methods in Molecular Biology, vol 1858. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8775-7_15

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  • DOI: https://doi.org/10.1007/978-1-4939-8775-7_15

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8774-0

  • Online ISBN: 978-1-4939-8775-7

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