期刊论文详细信息
Frontiers in Neuroscience
A Sextuple Knockout Cell Line System to Study the Differential Roles of CRY, PER, and NR1D in the Transcription-Translation Feedback Loop of the Circadian Clock
Yanyan Yang1  Aziz Sancar1  Yi-Ying Chiou2  Tzu-Ying Li2 
[1] Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, NC, United States;Graduate Institute of Biochemistry, National Chung Hsing University, Taichung City, Taiwan;
关键词: circadian clock;    TTFL;    Cryptochrome;    knockout;    serum shock;   
DOI  :  10.3389/fnins.2020.616802
来源: DOAJ
【 摘 要 】

The transcription-translation feedback loop (TTFL) is the core mechanism of the circadian rhythm. In mammalian cells, CLOCK-BMAL1 proteins activate the downstream genes by binding on the E-box sequence of the clock-controlled genes. Among these gene products, CRY1, CRY2, PER1, PER2, NR1D1, and NR1D2 can regulate the CLOCK-BMAL1-mediated transcription to form the feedback loop. However, the detailed mechanism of the TTFL is unclear because of the complicated inter-regulation of these proteins. Here, we generated a cell line lacking CRY1, CRY2, PER1, PER2, NR1D1, and NR1D2 (Cry/Per/Nr1d_KO) to study TTFL. We compared the Dbp transcription after serum-shock and dexamethasone-shock between Cry/Per/Nr1d_KO cells and cells expressing endogenous CRY (Per/Nr1d_KO) or NR1D (Cry/Per_KO). Furthermore, we found that CRY1-mediated repression of Dbp could persist more than 24 h in the absence of other proteins in the negative limb of the TTFL. Our Cry/Per/Nr1d_KO cells is a suitable system for the studying of differential roles of CRY, PER, and NR1D in the TTFL.

【 授权许可】

Unknown   

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