title: Chitosan/interfering RNA nanoparticle mediated gene silencing in disease vector mosquito larvae.
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.
authors: Division of Biology, Kansas State University.
authors: Xin
authors: X
authors: Zhang
authors: Department of Medical and Molecular Genetics, Indiana University School of Medicine; Eck Institute for Global Health, University of Notre Dame.
authors: Keshava
authors: K
authors: Mysore
authors: Eck Institute for Global Health, University of Notre Dame; Department of Biological Sciences, University of Notre Dame.
authors: Ellen
authors: E
authors: Flannery
authors: Division of Biology, Kansas State University.
authors: Kristin
authors: K
authors: Michel
authors: Eck Institute for Global Health, University of Notre Dame; Department of Biological Sciences, University of Notre Dame.
authors: David W
authors: DW
authors: Severson
authors: Department of Entomology, Kansas State University.
authors: Kun Yan
authors: KY
authors: Zhu
authors: Department of Medical and Molecular Genetics, Indiana University School of Medicine; Eck Institute for Global Health, University of Notre Dame; Department of Biological Sciences, University of Notre Dame; Molly.A.Scheel.2@nd.edu.
authors: Molly
authors: M
authors: Duman-Scheel
mesh_terms: D000330:Aedes / Q000235:genetics*; D000818:Animals; D000852:Anopheles / Q000235:genetics*; D048271:Chitosan / Q000008:administration & dosage / Q000737:chemistry; D005260:Female; D055785:Gene Knockdown Techniques / Q000379:methods*; D020868:Gene Silencing; D007303:Insect Vectors / Q000235:genetics*; D007814:Larva; D009032:Mosquito Control / Q000379:methods*; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry; D011679:Pupa; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics*
chemical_list: D012330:RNA, Double-Stranded; D034741:RNA, Small Interfering; D048271:Chitosan