Targeted gene expression in the transgenic Aedes aegypti using the binary Gal4-UAS system

• Published in: Insect biochemistry and molecular biology 2011-08, Vol.41 (8), p.637-644
• Main Authors: Kokoza, Vladimir A, Raikhel, Alexander S
• Format: Article
• Language: English
• Subjects: 5' Flanking Region; Aedes - drug effects; Aedes - genetics; Aedes - metabolism; Aedes aegypti; Animals; Animals, Genetically Modified; blood meal; ecdysterone; Ecdysterone - pharmacology; fat body; Fat Body - drug effects; Fat Body - metabolism; Female; females; gene expression; Gene Expression - drug effects; gene expression regulation; genes; Genes, Reporter; Genetic Engineering - methods; Genetic transformation; Green Fluorescent Proteins - analysis; hormonal regulation; hybrids; Insect Proteins - genetics; Insect Proteins - metabolism; organ culture; Organ Culture Techniques; PiggyBac transposable element; Plasmids; Promoter Regions, Genetic; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - analysis; Transcriptional Activation - drug effects; Transformation, Genetic; vitellogenin; Vitellogenin promoter; Vitellogenins - genetics; Vitellogenins - metabolism
• ISSN: 0965-1748; 1879-0240
• DOI: 10.1016/j.ibmb.2011.04.004

In this study, we report the establishment of the binary Gal4/UAS system for the yellow fever mosquito Aedes aegypti. We utilized the 1.8-kb 5' upstream region of the vitellogenin gene (Vg) to genetically engineer mosquito lines with the Vg-Gal4 activator and established UAS-EGFP responder transgenic mosquito lines to evaluate the binary Gal4/UAS system. The results show that the Vg-Gal4 driver leads to a high level of tissue-, stage- and sex-specific expression of the EGFP reporter in the fat body of Vg-Gal4/UAS-EGFP hybrids after blood-meal activation. In addition, the applicability of this system to study hormonal regulation of gene expression was demonstrated in in vitro organ culture experiments in which the EGFP reporter was highly activated in isolated fat bodies of previtellogenic Vg-Gal4/UAS-EGFP females incubated in the presence of 20-hydroxyecdysone (20E). Hence, this study has opened the door for further refinement of genetic tools in mosquitoes.