RNAi Trigger Delivery into Anopheles gambiae Pupae

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dc.contributor.author Regna, K.
dc.contributor.author Harrison, R. M.
dc.contributor.author Heyse, S. A.
dc.contributor.author Chiles, T. C.
dc.contributor.author Michel, Kristin
dc.contributor.author Muskavitch, M. A. T.
dc.date.accessioned 2016-09-20T14:53:44Z
dc.date.available 2016-09-20T14:53:44Z
dc.identifier.uri http://hdl.handle.net/2097/33955
dc.description Citation: Regna, K., Harrison, R. M., Heyse, S. A., Chiles, T. C., Michel, K., & Muskavitch, M. A. T. (2016). RNAi Trigger Delivery into Anopheles gambiae Pupae. Jove-Journal of Visualized Experiments(109), 9. doi:10.3791/53738
dc.description.abstract RNA interference (RNAi), a naturally occurring phenomenon in eukaryotic organisms, is an extremely valuable tool that can be utilized in the laboratory for functional genomic studies. The ability to knockdown individual genes selectively via this reverse genetic technique has allowed many researchers to rapidly uncover the biological roles of numerous genes within many organisms, by evaluation of loss-of-function phenotypes. In the major human malaria vector Anopheles gambiae, the predominant method used to reduce the function of targeted genes involves injection of double-stranded (dsRNA) into the hemocoel of the adult mosquito. While this method has been successful, gene knockdown in adults excludes the functional assessment of genes that are expressed and potentially play roles during pre-adult stages, as well as genes that are expressed in limited numbers of cells in adult mosquitoes. We describe a method for the injection of Serine Protease Inhibitor 2 (SRPN2) dsRNA during the early pupal stage and validate SRPN2 protein knockdown by observing decreased target protein levels and the formation of melanotic pseudo-tumors in SRPN2 knockdown adult mosquitoes. This evident phenotype has been described previously for adult stage knockdown of SRPN2 function, and we have recapitulated this adult phenotype by SRPN2 knockdown initiated during pupal development. When used in conjunction with a dye-labeled dsRNA solution, this technique enables easy visualization by simple light microscopy of injection quality and distribution of dsRNA in the hemocoel.
dc.relation.uri https://doi.org/10.3791/53738
dc.rights Attribution-NonCommercial-NoDerivs 3.0 United States (CC BY-NC-ND 3.0 US)
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/3.0/us/
dc.subject Infection
dc.subject Issue 109
dc.subject Mosquito
dc.subject Pupa
dc.subject Malaria
dc.subject Gambiae
dc.title RNAi Trigger Delivery into Anopheles gambiae Pupae
dc.type Article
dc.date.published 2016
dc.citation.doi 10.3791/53738
dc.citation.issn 1940-087X
dc.citation.issue 109
dc.citation.jtitle Jove-Journal of Visualized Experiments
dc.citation.spage 9
dc.contributor.authoreid kmichel

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Attribution-NonCommercial-NoDerivs 3.0 United States (CC BY-NC-ND 3.0 US) Except where otherwise noted, the use of this item is bound by the following: Attribution-NonCommercial-NoDerivs 3.0 United States (CC BY-NC-ND 3.0 US)

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