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 Scheel

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

LanguageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages213-231
Number of pages19
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1858
ISSN (Print)1064-3745

Fingerprint

Gene Silencing
Culicidae
Small Interfering RNA
Larva
Yeasts
Tablets
Saccharomyces cerevisiae
Anopheles gambiae
Disease Vectors
Dengue Virus
Genetic Engineering
Arthropods
Aedes
RNA Interference
Malaria
Eating
Genome
Research
Genes

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Mysore, K., Hapairai, L. K., Wei, N., Realey, J. S., Scheel, N. D., Severson, D. W., & Scheel, M. (2019). Preparation and use of a yeast shRNA delivery system for gene silencing in mosquito larvae. In Methods in Molecular Biology (pp. 213-231). (Methods in Molecular Biology; Vol. 1858). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8775-7_15

Preparation and use of a yeast shRNA delivery system for gene silencing in mosquito larvae. / Mysore, Keshava; Hapairai, Limb K.; Wei, Na; Realey, Jacob S.; Scheel, Nicholas D.; Severson, David W.; Scheel, Molly.

Methods in Molecular Biology. Humana Press Inc., 2019. p. 213-231 (Methods in Molecular Biology; Vol. 1858).

Research output: Chapter in Book/Report/Conference proceedingChapter

Mysore, K, Hapairai, LK, Wei, N, Realey, JS, Scheel, ND, Severson, DW & Scheel, M 2019, Preparation and use of a yeast shRNA delivery system for gene silencing in mosquito larvae. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1858, Humana Press Inc., pp. 213-231. https://doi.org/10.1007/978-1-4939-8775-7_15
Mysore K, Hapairai LK, Wei N, Realey JS, Scheel ND, Severson DW et al. Preparation and use of a yeast shRNA delivery system for gene silencing in mosquito larvae. In Methods in Molecular Biology. Humana Press Inc. 2019. p. 213-231. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-8775-7_15
Mysore, Keshava ; Hapairai, Limb K. ; Wei, Na ; Realey, Jacob S. ; Scheel, Nicholas D. ; Severson, David W. ; Scheel, Molly. / Preparation and use of a yeast shRNA delivery system for gene silencing in mosquito larvae. Methods in Molecular Biology. Humana Press Inc., 2019. pp. 213-231 (Methods in Molecular Biology).
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