RRC ID 54849
Author Zhang W, Takahara T, Achiha T, Shibata H, Maki M.
Title Cellular Ca2+-Responding Nanoluciferase Reporter Gene System Directed by Tandemly Repeated Pseudo-palindromic NFAT-Response Elements.
Journal Methods Mol Biol
Abstract Luciferase reporter gene systems based on the NFAT-response element (RE) have been used to monitor intracellular Ca2+ elevation. However, Ca2+ mobilization agent (e.g., ionomycin) alone is not adequate to activate the currently often employed reporter gene that contains the NFAT-RE found in the IL2 promoter. In addition to activation of NFAT through the Ca2+-calmodulin/calcineurin pathway, activation of AP-1 as a partner transcription factor is essential for the IL2-based NFAT-RE system. Here, we describe a detailed method for the recently developed new reporter gene system containing the NFAT-RE from the IL8 promoter. This system enables us to monitor endpoint effects of Ca2+-mobilizing agonists independent of AP-1 activation.
Volume 1929
Pages 95-109
Published 2019-1-1
DOI 10.1007/978-1-4939-9030-6_7
PMID 30710269
MeSH Calcineurin / genetics Calcium / analysis* Calmodulin / genetics Gene Expression Regulation Genes, Reporter* HEK293 Cells Humans Interleukin-2 / chemistry Interleukin-2 / genetics* NFATC Transcription Factors / metabolism Promoter Regions, Genetic Transcription Factor AP-1 / genetics Transcriptional Activation
Human and Animal Cells 293(RCB1637)