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Bidirectional role of orphan nuclear receptor RORα in clock gene transcriptions demonstrated by a novel reporter assay system
Author(s) -
Nakajima Yoshihiro,
Ikeda Masaaki,
Kimura Takuma,
Honma Sato,
Ohmiya Yoshihiro,
Honma Ken-ichi
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2004.03.083
Subject(s) - per1 , reporter gene , enhancer , promoter , neuron derived orphan receptor 1 , nuclear receptor , circadian clock , biology , orphan receptor , transcription factor , microbiology and biotechnology , clock , transcription (linguistics) , luciferases , gene , luciferase , genetics , chemistry , gene expression , transfection , linguistics , philosophy
Circadian rhythms are generated by an extremely complicated transcription–translation feedback loop. To precisely analyze the molecular mechanisms of the circadian clock, it is critical to monitor multiple gene expressions and/or interactions with their transcription factors simultaneously. We have developed a novel reporter assay system, the tricolor reporter in vitro assay system, which consists of green‐ and red‐emitting Phrixothrix luciferases as dual reporters and blue‐emitting Renilla luciferase as internal control. We have successfully employed this system in analyzing the effects of clock gene products on the enhancer elements of Per1 and Bmal1 promoters. The results indicate that the orphan nuclear receptor RORα regulates bidirectionally Bmal1 (positively) and Per1 (negatively) transcriptions simultaneously.