Browsing by Author "Hapairai, Limb"

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    Oral RNAi for Gene Silencing in Mosquitoes: From the Bench to the Field
    (Cold Spring Harbor Laboratory Press, 2022-07-12) Mysore, Keshava; Hapairai, Limb; Realey, Jacob S.; Sun, Longhua; Roethele, Joseph B.; Duman-Scheel, Molly; Medical and Molecular Genetics, School of Medicine
    RNA interference (RNAi) has played a key role in the field of insect functional genomics, a discipline that has enhanced the study of developmental, evolutionary, physiological, and molecular biological phenomena in a wide variety of insects, including disease vector mosquitoes. Here we introduce a recently optimized RNAi procedure in which adult mosquitoes are fed with a colored sugar bait containing small interfering RNA (siRNA). This procedure effectively and economically leads to gene silencing, is technically straightforward, and has been successfully used to characterize a number of genes in adult mosquitoes. We also discuss how, in addition to laboratory applications, this oral RNAi procedure might one day be used in the field for controlling insect pests.
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    Sugar-Baited Delivery of Small Interfering RNA for Gene Silencing in Adult Mosquitoes
    (Cold Spring Harbor Laboratory, 2022-07-12) Mysore, Keshava; Hapairai, Limb; Realey, Jacob S.; Sun, Longhua; Roethele, Joseph B.; Duman-Scheel, Molly; Medical and Molecular Genetics, School of Medicine
    RNA interference (RNAi), an innate regulatory mechanism that is conserved across many eukaryotic species, has been harnessed for experimental gene silencing in many organisms, including mosquitoes. This protocol describes an optimized method for inducing RNAi in adult Aedes aegypti and Anopheles gambiae mosquitoes that involves feeding them a red-colored sugar bait containing small interfering RNA (siRNA). This oral delivery method is less physically disruptive than delivery by subcutaneous injection, and the use of siRNAs (in contrast to long dsRNAs) for RNAi enables the design of molecules that target conserved sites so that gene function can be studied in multiple species. After feeding, the behavioral and morbidity phenotypes that result from the suppression of target gene expression can then be analyzed.