A timer for analyzing temporally dynamic changes in transcription during differentiation in vivo.
A timer for analyzing temporally dynamic changes in transcription during differentiation in vivo.
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DOI：
10.1083/jcb.201711048
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发表时间：
2018-08-06
期刊：
影响因子：
--
通讯作者：
Ono M
中科院分区：
文献类型：
--
作者：
Bending D;Prieto Martín P;Paduraru A;Ducker C;Marzaganov E;Laviron M;Kitano S;Miyachi H;Crompton T;Ono M
bend等人建立了一个新的工具——Timer of cell kinetics and activity (Tocky)，揭示了细胞在体内激活和分化的时间动态。该工具分析细胞分化过程中分子过程的时间序列，并可以根据细胞在体内接收信号事件的频率对细胞进行分类。理解细胞分化的机制是具有挑战性的，因为分化是由驱动暂时动态过程的信号通路启动的，而这些过程很难在体内分析。我们建立了一个新的工具，细胞动力学和活性定时器（Tocky；或toki[日语时间]）。Tocky使用荧光Timer蛋白，它可以自发地将其发射光谱从蓝色转变为红色，并结合计算机算法来揭示体内分化的动态。利用T细胞受体（TCR）信号的转录靶点，我们建立了Nr4a3-Tocky来跟踪TCR信号的下游效应。Nr4a3-Tocky揭示了调节性T细胞（Treg）分化过程中事件的时间序列，并表明在Treg生成过程中持续的TCR信号发生。值得注意的是，自身免疫性炎症部位的抗原特异性T细胞也显示出持续的TCR信号。此外，通过生成Foxp3- tocky，我们揭示了Foxp3基因去甲基化的体内动态。因此，Tocky是细胞生物学家通过直接揭示体内动态过程来解决以前无法解决的问题的工具。
Bending et al. establish a new tool, Timer of cell kinetics and activity (Tocky), revealing the temporal dynamics of cellular activation and differentiation in vivo. The tool analyzes the temporal sequence of molecular processes during cellular differentiation and can classify cells based on the frequency they receive signaling events in vivo. Understanding the mechanisms of cellular differentiation is challenging because differentiation is initiated by signaling pathways that drive temporally dynamic processes, which are difficult to analyze in vivo. We establish a new tool, Timer of cell kinetics and activity (Tocky; or toki [time in Japanese]). Tocky uses the fluorescent Timer protein, which spontaneously shifts its emission spectrum from blue to red, in combination with computer algorithms to reveal the dynamics of differentiation in vivo. Using a transcriptional target of T cell receptor (TCR) signaling, we establish Nr4a3-Tocky to follow downstream effects of TCR signaling. Nr4a3-Tocky reveals the temporal sequence of events during regulatory T cell (Treg) differentiation and shows that persistent TCR signals occur during Treg generation. Remarkably, antigen-specific T cells at the site of autoimmune inflammation also show persistent TCR signaling. In addition, by generating Foxp3-Tocky, we reveal the in vivo dynamics of demethylation of the Foxp3 gene. Thus, Tocky is a tool for cell biologists to address previously inaccessible questions by directly revealing dynamic processes in vivo.
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DOI：
10.4049/jimmunol.1300408
发表时间：
2013-10-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Li L;Nishio J;van Maurik A;Mathis D;Benoist C
通讯作者：
Benoist C
影响因子：
4.4
作者：
Burchill, Matthew A.;Yang, Jianying;Farrar, Michael A.
通讯作者：
Farrar, Michael A.
影响因子：
3.3
作者：
O'Neill EJ;Day MJ;Wraith DC
通讯作者：
Wraith DC
影响因子：
32.4
作者：
Lio, Chan-Wang Joaquim;Hsieh, Chyi-Song
通讯作者：
Hsieh, Chyi-Song
影响因子：
16.6
作者：
Koechlein CS;Harris JR;Lee TK;Weeks J;Fox RG;Zimdahl B;Ito T;Blevins A;Jung SH;Chute JP;Chourasia A;Covert MW;Reya T
通讯作者：
Reya T