Chitosan/interfering RNA nanoparticle mediated gene silencing in disease vector mosquito larvae

Xin Zhang, Keshava Mysore, Ellen Flannery, Kristin Michel, David W. Severson, Kun Yan Zhu, Molly Duman-Scheel

Research output: Contribution to journalArticle

37 Scopus citations

Abstract

Vector mosquitoes inflict more human suffering than any other organism—and kill more than one million people each year. The mosquito genome projects facilitated research in new facets of mosquito biology, including functional genetic studies in the primary African malaria vector Anopheles gambiae and the dengue and yellow fever vector Aedes aegypti. RNA interference- (RNAi-) mediated gene silencing has been used to target genes of interest in both of these disease vector mosquito species. Here, we describe a procedure for preparation of chitosan/ interfering RNA nanoparticles that are combined with food and ingested by larvae. This technically straightforward, high-throughput, and relatively inexpensive methodology, which is compatible with long double stranded RNA (dsRNA) or small interfering RNA (siRNA) molecules, has been used for the successful knockdown of a number of different genes in A. gambiae and A. aegypti larvae. Following larval feedings, knockdown, which is verified through qRT-PCR or in situ hybridization, can persist at least through the late pupal stage. This methodology may be applicable to a wide variety of mosquito and other insect species, including agricultural pests, as well as other non-model organisms. In addition to its utility in the research laboratory, in the future, chitosan, an inexpensive, non-toxic and biodegradable polymer, could potentially be utilized in the field.

Original languageEnglish (US)
Article numbere52523
JournalJournal of Visualized Experiments
Volume2015
Issue number97
DOIs
StatePublished - Mar 25 2015

Keywords

  • Aedes aegypti
  • Anopheles gambiae
  • Development
  • Disease
  • DsRNA
  • Ingestion
  • Issue 97
  • Knockdown
  • Larvae
  • Molecular biology
  • Mosquito
  • RNA interference
  • SiRNA
  • Vector biology

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

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