title: Targeted gene expression in the transgenic Aedes aegypti using the binary Gal4-UAS system.
abstract: 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.
authors.affiliation: Department of Entomology and Institute for Integrative Genome Biology, University of California Riverside, Riverside, CA 92521, USA.
authors.forename: Vladimir A
authors.initials: VA
authors.lastname: Kokoza
authors.forename: Alexander S
authors.initials: AS
authors.lastname: Raikhel
mesh_terms: D024506:5' Flanking Region; D000330:Aedes / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D000818:Animals; D030801:Animals, Genetically Modified; D004441:Ecdysterone / Q000494:pharmacology*; D005216:Fat Body / Q000187:drug effects* / Q000378:metabolism; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter; D005818:Genetic Engineering / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000032:analysis; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D009924:Organ Culture Techniques; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015533:Transcriptional Activation / Q000187:drug effects*; D014170:Transformation, Genetic; D014819:Vitellogenins / Q000235:genetics* / Q000378:metabolism
chemical_list: D019476:Insect Proteins; D012333:RNA, Messenger; D014819:Vitellogenins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004441:Ecdysterone