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Using the GAL4-UAS System for Functional Genetics in Anopheles gambiae
Published on: April 15, 2021
Driving midgut-specific expression and secretion of a foreign protein in transgenic mosquitoes with AgAper1
E G Abraham1, M Donnelly-Doman, H Fujioka
1Department of Molecular Microbiology and Immunology, John Hopkins Bloomberg School of Public Health, Malaria Research Institute, Baltimore, MD 21205, USA.
Abstract:
The Anopheles gambiae adult peritrophic matrix protein 1 (AgAper1) regulatory elements were used to drive the expression of phospholipase A2 (PLA2), a protein known to disrupt malaria parasite development in mosquitoes. These AgAper1 regulatory elements were sufficient to promote the accumulation of PLA2 in midgut epithelial cells before a blood meal and its release into the lumen upon blood ingestion. Plasmodium berghei oocyst formation was reduced by approximately 80% (74-91% range) in transgenic mosquitoes. Blood-seeking behaviour and survival of AgAper1-PLA2 transgenic mosquitoes were comparable to sibling wild-type mosquitoes, while fertility was substantially lower. Ultrastructural studies suggest that decreased fitness is a consequence of internal damage to midgut epithelial cells.
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