Dual-controlled optogenetic system for the rapid down-regulation of protein levels in mammalian cells.
Dual-controlled optogenetic system for the rapid down-regulation of protein levels in mammalian cells.
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DOI：
10.1038/s41598-018-32929-7
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发表时间：
2018-10-09
影响因子：
4.6
通讯作者：
Zurbriggen MD
中科院分区：
文献类型：
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作者：
Baaske J;Gonschorek P;Engesser R;Dominguez-Monedero A;Raute K;Fischbach P;Müller K;Cachat E;Schamel WWA;Minguet S;Davies JA;Timmer J;Weber W;Zurbriggen MD
光遗传开关是研究细胞过程的新兴分子工具，因为它们比传统的基于化学的开关提供更高的时空和定量精度。光可控基因表达系统旨在上调蛋白质表达水平，同时表现出优于基于化学的对应物的性能。然而，以类似效率降低蛋白质水平的系统却落后了。在这里，我们提出了一种新的双组分，蓝光响应光遗传开关（' blue -OFF '），它可以在光照下快速定量地下调蛋白质。blue - off将首个光响应抑制因子krabb - el222与蛋白质降解模块B-LID（蓝光诱导降解结构域）结合在一起，以单一波长同时控制基因表达和蛋白质稳定性。因此，Blue-OFF在控制蛋白质水平方面优于目前的光遗传系统。该系统是由一个数学模型来描述的，该模型有助于选择实验条件，如光强度和照明制度，以获得所需的结果。这种方法代表了双控光遗传系统的进步，其中多个光传感模块协同工作。正如在哺乳动物细胞培养中控制细胞凋亡的例子，该方法为基础研究和应用（如组织工程）开辟了新的视角。
Optogenetic switches are emerging molecular tools for studying cellular processes as they offer higher spatiotemporal and quantitative precision than classical, chemical-based switches. Light-controllable gene expression systems designed to upregulate protein expression levels meanwhile show performances superior to their chemical-based counterparts. However, systems to reduce protein levels with similar efficiency are lagging behind. Here, we present a novel two-component, blue light-responsive optogenetic OFF switch (‘Blue-OFF’), which enables a rapid and quantitative down-regulation of a protein upon illumination. Blue-OFF combines the first light responsive repressor KRAB-EL222 with the protein degradation module B-LID (blue light-inducible degradation domain) to simultaneously control gene expression and protein stability with a single wavelength. Blue-OFF thus outperforms current optogenetic systems for controlling protein levels. The system is described by a mathematical model which aids in the choice of experimental conditions such as light intensity and illumination regime to obtain the desired outcome. This approach represents an advancement of dual-controlled optogenetic systems in which multiple photosensory modules operate synergistically. As exemplified here for the control of apoptosis in mammalian cell culture, the approach opens up novel perspectives in fundamental research and applications such as tissue engineering.
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影响因子：
14.9
作者：
Müller K;Engesser R;Schulz S;Steinberg T;Tomakidi P;Weber CC;Ulm R;Timmer J;Zurbriggen MD;Weber W
通讯作者：
Weber W
影响因子：
1.5
作者：
Chu, Yuanyuan;Senghaas, Niklas;Kuehn, Ralf
通讯作者：
Kuehn, Ralf
影响因子：
4.7
作者：
Kolar, Katja;Knobloch, Christian;Weber, Wilfried
通讯作者：
Weber, Wilfried
影响因子：
56.9
作者：
Drab, M;Verkade, P;Kurzchalia, TV
通讯作者：
Kurzchalia, TV
DOI：
10.1038/nrmicro2622
发表时间：
2011-08-08
期刊：
Nature reviews. Microbiology
影响因子：
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作者：
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