A synthetic BRET-based optogenetic device for pulsatile transgene expression enabling glucose homeostasis in mice.
A synthetic BRET-based optogenetic device for pulsatile transgene expression enabling glucose homeostasis in mice.
复制标题
一种基于 BRET 的合成光遗传学装置，用于脉冲式转基因表达，使小鼠体内的葡萄糖稳态得以实现。
DOI：
10.1038/s41467-021-20913-1
复制
发表时间：
2021-01-27
影响因子：
16.6
通讯作者：
Yang Y
中科院分区：
文献类型：
--
作者：
Li T;Chen X;Qian Y;Shao J;Li X;Liu S;Zhu L;Zhao Y;Ye H;Yang Y
基因电路中的脉冲细胞动力学已被证明为细胞在应激反应、信号传导和发育中提供关键能力。尽管在过去的几年中取得了令人着迷的发现，大多数脉冲系统的机制和功能仍然不清楚，关键的挑战之一是缺乏一种技术，允许在体外和体内的转基因表达的脉冲调节。在这里，我们描述了一种基于发光转录因子（称为luminGAVPO）的合成BRET转基因表达（LuminON）系统的开发，该系统通过将NanoLuc荧光素酶融合到光可切换转录因子GAVPO。luminGAVPO允许在哺乳动物细胞和小鼠中通过化学遗传学和光遗传学方法脉冲和定量激活转基因表达。转基因表达的脉冲幅度和持续时间都是通过调节丙氨嗪的量高度可调的。我们进一步证明了LuminON介导的1型糖尿病小鼠的血糖稳态。我们相信，基于BRET的LuminON系统与转基因表达的脉动动力学提供了一个高度敏感的工具，在生物系统中的精确操作，具有强大的潜力，在未来的各种基础生物学研究和基因和细胞为基础的精确治疗的应用。基因电路中的脉冲细胞动力学已被证明在不同的细胞活动中为细胞提供关键能力。在这里，作者展示了一种基于BRET的合成转基因表达系统，该系统允许活细胞和体内基因表达的脉冲和定量激活。
Pulsing cellular dynamics in genetic circuits have been shown to provide critical capabilities to cells in stress response, signaling and development. Despite the fascinating discoveries made in the past few years, the mechanisms and functional capabilities of most pulsing systems remain unclear, and one of the critical challenges is the lack of a technology that allows pulsatile regulation of transgene expression both in vitro and in vivo. Here, we describe the development of a synthetic BRET-based transgene expression (LuminON) system based on a luminescent transcription factor, termed luminGAVPO, by fusing NanoLuc luciferase to the light-switchable transcription factor GAVPO. luminGAVPO allows pulsatile and quantitative activation of transgene expression via both chemogenetic and optogenetic approaches in mammalian cells and mice. Both the pulse amplitude and duration of transgene expression are highly tunable via adjustment of the amount of furimazine. We further demonstrated LuminON-mediated blood-glucose homeostasis in type 1 diabetic mice. We believe that the BRET-based LuminON system with the pulsatile dynamics of transgene expression provides a highly sensitive tool for precise manipulation in biological systems that has strong potential for application in diverse basic biological studies and gene- and cell-based precision therapies in the future. Pulsing cellular dynamics in genetic circuits have been shown to provide critical capabilities to cells in diverse cellular activities. Here the authors show a synthetic BRET-based transgene expression system that allows pulsatile and quantitative activation of gene expression both in live cells and in vivo.
登录
查看更多内容
影响因子：
48
作者：
Kennedy, Matthew J.;Hughes, Robert M.;Peteya, Leslie A.;Schwartz, Joel W.;Ehlers, Michael D.;Tucker, Chandra L.
通讯作者：
Tucker, Chandra L.
影响因子：
5.4
作者：
Castle, Jessica R.;DeVries, J. Hans;Kovatchev, Boris
通讯作者：
Kovatchev, Boris
DOI：
10.1073/pnas.1510899113
发表时间：
2016-01-19
影响因子：
11.1
作者：
Berglund, Ken;Clissold, Kara;Hochgeschwender, Ute
通讯作者：
Hochgeschwender, Ute
影响因子：
3.3
作者：
Fu, Zhuo;Gilbert, Elizabeth R.;Liu, Dongmin
通讯作者：
Liu, Dongmin
影响因子：
5.1
作者：
Liu, F;Song, YK;Liu, D
通讯作者：
Liu, D