Targeted modification of the Per2 clock gene alters circadian function in mPer2luciferase (mPer2Luc) mice.
Targeted modification of the Per2 clock gene alters circadian function in mPer2luciferase (mPer2Luc) mice.
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DOI：
10.1371/journal.pcbi.1008987
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发表时间：
2021-05
影响因子：
4.3
通讯作者：
Sumová A
中科院分区：
文献类型：
--
作者：
Ralph MR;Shi SQ;Johnson CH;Houdek P;Shrestha TC;Crosby P;O'Neill JS;Sládek M;Stinchcombe AR;Sumová A
修饰mPer 2Luc报告小鼠中的Per 2时钟基因显著改变昼夜节律功能。在恒定黑暗中的行为周期延长，并在恒定光照下分解为两个不同的组成部分。节律表现出增加双峰，增强相位重置光脉冲，并改变夹带预定喂养。机械数学模型预测，增强蛋白质与修饰mPER 2 C-末端的相互作用，结合SCN亚区之间的差异时钟调节，可以通过增加Per 2转录和蛋白质稳定性来解释对昼夜节律行为的影响。PER 2：：LUC对CLOCK：BMAL 1 E-box活性的抑制作用大于PER 2。mPer 2Luc在Per 2的外显子23中携带72 bp缺失，并保留影响节律幅度但不影响周期的新霉素抗性盒。结果表明，mPer 2 Luc充当昼夜节律钟突变，表明需要详细评估C末端标签在转基因动物模型中的潜在影响。经修饰以表达生物发光信号的工程化基因可用于通过真实的时间反映过程的实际状态来监测细胞过程。对于昼夜节律研究，广泛使用的报告模型（mPer 2Luc）在时钟基因Period 2（Per 2）的控制下表达萤火虫荧光素酶，其编码产生生物发光节律的融合蛋白（PER 2：：LUCIFERASE）。我们提出的证据表明，工程蛋白质产生广泛的变化，在小鼠的标志性昼夜行为模式。证据来自对mPer 2Luc小鼠在恒定黑暗和恒定光照下的昼夜节律行为的长期记录的分析，以及对光脉冲的反应和对有限食物可用性的夹带。通过将视交叉上核中昼夜节律产生的成熟数学模型与核内已知的功能异质性相结合，我们概括了WT和mPer 2Luc小鼠中观察到的运动行为模式。从模型预测的PER 2蛋白稳定性的功能改变通过PER 2：：LUC相对于WT PER 2对CLOCK：BMAL 1转录激活的增加的抑制强度来证实。因此，mPer 2Luc是一种时钟突变，为哺乳动物昼夜节律系统的功能性神经解剖学和Per 2调节在节律产生中的至关重要性提供了新的见解。
Modification of the Per2 clock gene in mPer2Luc reporter mice significantly alters circadian function. Behavioral period in constant dark is lengthened, and dissociates into two distinct components in constant light. Rhythms exhibit increased bimodality, enhanced phase resetting to light pulses, and altered entrainment to scheduled feeding. Mechanistic mathematical modelling predicts that enhanced protein interactions with the modified mPER2 C-terminus, combined with differential clock regulation among SCN subregions, can account for effects on circadian behavior via increased Per2 transcript and protein stability. PER2::LUC produces greater suppression of CLOCK:BMAL1 E-box activity than PER2. mPer2Luc carries a 72 bp deletion in exon 23 of Per2, and retains a neomycin resistance cassette that affects rhythm amplitude but not period. The results show that mPer2Luc acts as a circadian clock mutation illustrating a need for detailed assessment of potential impacts of c-terminal tags in genetically modified animal models. Engineered genes that are modified to express bioluminescent signals can be used to monitor cellular processes by reflecting the actual state of the process in real time. For circadian rhythm studies, a widely used reporter model (mPer2Luc) expresses firefly luciferase under the control of the clock gene Period 2 (Per2), encoding a fusion protein (PER2::LUCIFERASE) which produces rhythms of bioluminescence. We present evidence that the engineered protein produces extensive changes in iconic circadian behavioral patterns in mice. The evidence comes from analysis of long-term recordings of circadian behavior in mPer2Luc mice in constant darkness and constant light, together with responsiveness to light pulses and entrainment to restricted food availability. By integrating a well-established mathematical model of circadian rhythm generation in the suprachiasmatic nuclei with the known functional heterogeneity within the nucleus, we recapitulated the observed locomotor behavioral patterns in WT and mPer2Luc mice. Functional alterations to the stability of PER2 protein predicted from the model were substantiated by increased repression strength of PER2::LUC over WT PER2 on CLOCK:BMAL1 transcriptional activation. Therefore, mPer2Luc is a clock mutation that provides new insights into the functional neuroanatomy of the mammalian circadian system and the critical importance of Per2 regulation in rhythm generation.
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DOI：
10.1089/153623103322637698
发表时间：
2003-12-01
期刊：
OMICS A Journal of Integrative Biology
影响因子：
--
作者：
Forger, Daniel B.;Dean, Dennis A., II;Weaver, David R.
通讯作者：
Weaver, David R.
DOI：
10.1073/pnas.1612917113
发表时间：
2016-10-11
影响因子：
11.1
作者：
Chiou, Yi-Ying;Yang, Yanyan;Sancar, Aziz
通讯作者：
Sancar, Aziz
影响因子：
3.7
作者：
Etchegaray, Jean-Pierre;Yu, Elizabeth A.;Weaver, David R.
通讯作者：
Weaver, David R.
影响因子：
4.3
作者：
Diekman CO;Belle MD;Irwin RP;Allen CN;Piggins HD;Forger DB
通讯作者：
Forger DB
影响因子：
9.2
作者：
de la Iglesia, HO;Cambras, T;Díez-Noguera, A
通讯作者：
Díez-Noguera, A