A luciferase reporter assay for ecdysone agonists using HEK293T cells

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...

Full description

Saved in:
Bibliographic Details
Published inBioscience, biotechnology, and biochemistry Vol. 86; no. 11; pp. 1490 - 1496
Main Authors Yokoi, Taiyo, Nabe, Taku, Ishizuka, Chiharu, Hayashi, Ken'ichiro, Ito-Harashima, Sayoko, Yagi, Takashi, Nakagawa, Yoshiaki, Miyagawa, Hisashi
Format Journal Article
LanguageEnglish
Published 20.10.2022
Online AccessGet full text

Cover

More Information
Summary: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.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:1347-6947
1347-6947
DOI:10.1093/bbb/zbac139