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

  • Keshava Mysore
  • Limb K. Hapairai
  • Na Wei
  • Jacob S. Realey
  • Nicholas D. Scheel
  • David W. Severson
  • Molly Duman-ScheelEmail author
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

Aedes aegypti Anopheles gambiae Saccharomyces cerevisiae RNAi Dengue Zika Malaria Vector Development Insect 

Notes

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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Keshava Mysore
    • 1
    • 2
  • Limb K. Hapairai
    • 1
    • 2
  • Na Wei
    • 2
    • 3
  • Jacob S. Realey
    • 1
    • 2
  • Nicholas D. Scheel
    • 2
    • 4
  • David W. Severson
    • 1
    • 2
    • 4
  • Molly Duman-Scheel
    • 1
    • 2
    • 4
    Email author
  1. 1.Department of Medical and Molecular GeneticsIndiana University School of MedicineSouth BendUSA
  2. 2.Eck Institute for Global HealthUniversity of Notre DameNotre DameUSA
  3. 3.Department of Civil and Environmental Engineering and Earth SciencesUniversity of Notre DameNotre DameUSA
  4. 4.Department of Biological SciencesUniversity of Notre DameNotre DameUSA

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