Improved transient silencing of gene expression in the mosquito female Aedes aegypti

McFarlane, M. , Laureti, M., Levée, T., Terry, S., Kohl, A. and Pondeville, E. (2021) Improved transient silencing of gene expression in the mosquito female Aedes aegypti. Insect Molecular Biology, 30(3), pp. 355-365. (doi: 10.1111/imb.12700) (PMID:33715239)

Abstract

Gene silencing using RNA interference (RNAi) has become a widely used genetic technique to study gene function in many organisms. In insects, this technique is often applied through the delivery of dsRNA. In the adult female Aedes aegypti, a main vector of human‐infecting arboviruses, efficiency of gene silencing following dsRNA injection varies greatly according to targeted genes. Difficult knockdowns using dsRNA can thus hamper gene function analysis. Here, by analysing silencing of three different genes in female Ae. aegypti (p400, ago2 and E75), we show that gene silencing can indeed be dsRNA sequence dependent but different efficiencies do not correlate with dsRNA length. Our findings suggest that silencing is likely also gene dependent, probably due to gene‐specific tissue expression and/or feedback mechanisms. We demonstrate that use of high doses of dsRNA can improve knockdown efficiency, and injection of a transfection reagent along with dsRNA reduces the variability in efficiency between replicates. Finally, we show that gene silencing cannot be achieved using siRNA injection in Ae. aegypti adult females. Overall, this work should help future gene function analyses using RNAi in adult females Ae. aegypti, leading towards a better understanding of physiological and infectious processes.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Mcdonald, Dr Melanie and Terry, Mrs Sandra and Pondeville, Dr Emilie and Kohl, Professor Alain

Creator Roles

McFarlane, M.: Data curation, Formal analysis, Investigation, Methodology, Supervision, Validation, Visualization, Writing – original draft, Writing – review and editing
Terry, S.: Resources
Kohl, A.: Funding acquisition, Writing – review and editing
Pondeville, E.: Conceptualization, Data curation, Formal analysis, Funding acquisition, Methodology, Project administration, Supervision, Validation, Writing – original draft, Writing – review and editing

• Authors: McFarlane, M., Laureti, M., Levée, T., Terry, S., Kohl, A., and Pondeville, E.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity; College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research
• Journal Name: Insect Molecular Biology
• Publisher: Wiley
• ISSN: 0962-1075
• ISSN (Online): 1365-2583
• Published Online: 14 March 2021
• Copyright Holders: Copyright © 2021 The Authors
• First Published: First published in Insect Molecular Biology 30(3): 355-365
• Publisher Policy: Reproduced under a Creative Commons License

Project Code	Award No	Project Name	Principal Investigator	Funder's Name	Funder Ref	Lead Dept
				Medical Research Council (MRC)	MC_UU_12014/8