RNAi-mediated gene knockdown and in vivo diuresis assay in adult female Aedes aegypti mosquitoes
Autor(es): Drake Lisa L,Price David P,Aguirre Sarah E,Hansen Immo A
Resumo: 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.
Palavras-Chave: Genetics; Issue 65; Molecular Biology; Infection; Diuresis; Malpighian tubules; RNA interference; Aedes aegypti; Aquaporin
Imprenta: Journal of visualized experiments : JoVE, v. 65, p. e3479, 2012
Identificador do objeto digital: 10.3791/3479
Descritores: Aedes aegypti - Molecular Structure ; Aedes aegypti - Pathogenesis ; Aedes aegypti - RNA
Data de publicação: 2012
