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

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 tran...

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Bibliographic Details
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
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Summary: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.
ISSN:0965-1748
1879-0240
DOI:10.1016/j.ibmb.2011.04.004