title: RNAi-mediated gene knockdown and in vivo diuresis assay in adult female Aedes aegypti mosquitoes.
abstract: This video protocol demonstrates an effective technique to knockdown a particular gene in an insect and conduct a novel bioassay to measure excretion rate. This method can be used to obtain a better understanding of the process of diuresis in insects and is especially useful in the study of diuresis in blood-feeding arthropods that are able to take up huge amounts of liquid in a single blood meal. This RNAi-mediated gene knockdown combined with an in vivo diuresis assay was developed by the Hansen lab to study the effects of RNAi-mediated knockdown of aquaporin genes on Aedes aegypti mosquito diuresis. The protocol is setup in two parts: the first demonstration illustrates how to construct a simple mosquito injection device and how to prepare and inject dsRNA into the thorax of mosquitoes for RNAi-mediated gene knockdown. The second demonstration illustrates how to determine excretion rates in mosquitoes using an in vivo bioassay.
authors: Biology Department, New Mexico State University, USA.
authors: Lisa L
authors: LL
authors: Drake
authors: David P
authors: DP
authors: Price
authors: Sarah E
authors: SE
authors: Aguirre
authors: Immo A
authors: IA
authors: Hansen
mesh_terms: D000330:Aedes / Q000235:genetics*; D000818:Animals; D004231:Diuresis / Q000235:genetics; D005260:Female; D055785:Gene Knockdown Techniques / Q000379:methods*; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000008:administration & dosage* / Q000235:genetics*
chemical_list: D012330:RNA, Double-Stranded