We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
A luciferase reporter assay for ecdysone agonists using HEK293T cells.
- Authors
Taiyo Yokoi; Taku Nabe; Chiharu Ishizuka; Ken'ichiro Hayashi; Sayoko Ito-Harashima; Takashi Yagi; Yoshiaki Nakagawa; Hisashi Miyagawa
- Abstract
Ecdysone agonists are a class of insecticides that activate the ecdysone receptor (EcR) heterodimerized with the ultraspiracle (USP). Here, we report a new luciferase reporter assay for ecdysone agonists. The assay employs mammalian HEK293T cells transiently transfected with the EcR and USP genes of Chilo suppressalis, along with the taiman (Tai) gene of Drosophila melanogaster that encodes a steroid receptor coactivator. This assay system gave results consistent with those of radioligand binding assays and showed sensitivity superior to that of the existing in vitro methods. In addition, use of the heterologous host cells precludes perturbation from intrinsic players of the ecdysone signaling, which is a potential drawback of insect cell-based methods. This reporter system is suitable for detailed structure-activity analysis of ecdysone agonists and will serve as a valuable tool for the rational design of novel insect growth regulators.
- Subjects
ECDYSONE; INSECT growth regulators; CHILO suppressalis; STEROID receptors; RADIOLIGAND assay; DROSOPHILA melanogaster; LUCIFERASES
- Publication
Bioscience, Biotechnology & Biochemistry, 2022, Vol 86, Issue 11, p1490
- ISSN
0916-8451
- Publication type
Article
- DOI
10.1093/bbb/zbac139