Beyer Hannes M, Juillot Samuel, Herbst Kathrin, Samodelov Sophia L, Müller Konrad, Schamel Wolfgang W, Römer Winfried, Schäfer Eberhard, Nagy Ferenc, Strähle Uwe, Weber Wilfried, Zurbriggen Matias D
Faculty of Biology, University of Freiburg , Schänzlestrasse 1, 79104 Freiburg, Germany.
BIOSS - Centre for Biological Signalling Studies, University of Freiburg , Schänzlestrasse 18, 79104 Freiburg, Germany.
ACS Synth Biol. 2015 Sep 18;4(9):951-8. doi: 10.1021/acssynbio.5b00004. Epub 2015 Mar 30.
Protein trafficking in and out of the nucleus represents a key step in controlling cell fate and function. Here we report the development of a red light-inducible and far-red light-reversible synthetic system for controlling nuclear localization of proteins in mammalian cells and zebrafish. First, we synthetically reconstructed and validated the red light-dependent Arabidopsis phytochrome B nuclear import mediated by phytochrome-interacting factor 3 in a nonplant environment and support current hypotheses on the import mechanism in planta. On the basis of this principle we next regulated nuclear import and activity of target proteins by the spatiotemporal projection of light patterns. A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression. These data demonstrate the first in vivo application of a plant phytochrome-based optogenetic tool in vertebrates and expand the repertoire of available light-regulated molecular devices.
蛋白质进出细胞核的运输是控制细胞命运和功能的关键步骤。在此，我们报告了一种用于控制哺乳动物细胞和斑马鱼中蛋白质核定位的红光诱导且远红光可逆的合成系统的开发。首先，我们在非植物环境中合成重建并验证了由光敏色素相互作用因子3介导的红光依赖性拟南芥光敏色素B核输入，并支持了目前关于植物中输入机制的假说。基于这一原理，我们接下来通过光模式的时空投射来调节靶蛋白的核输入和活性。一种合成转录因子被转运到哺乳动物细胞和斑马鱼的细胞核中以驱动转基因表达。这些数据证明了基于植物光敏色素的光遗传学工具在脊椎动物中的首次体内应用，并扩展了可用的光调节分子装置的种类。
