Müller Konrad, Engesser Raphael, Timmer Jens, Zurbriggen Matias D, Weber Wilfried
Faculty of Biology, University of Freiburg , Schänzlestrasse 1, 79104 Freiburg, Germany.
ACS Synth Biol. 2014 Nov 21;3(11):796-801. doi: 10.1021/sb500305v. Epub 2014 Oct 28.
Optogenetic gene switches allow gene expression control at an unprecedented spatiotemporal resolution. Recently, light-responsive transgene expression systems that are activated by UV-B, blue, or red light have been developed. These systems perform well on their own, but their integration into genetic networks has been hampered by the overlapping absorbance spectra of the photoreceptors. We identified a lack of orthogonality between UV-B and blue light-controlled gene expression as the bottleneck and employed a model-based approach that identified the need for a blue light-responsive gene switch that is insensitive to low-intensity light. Based on this prediction, we developed a blue light-responsive and rapidly reversible expression system. Finally, we employed this expression system to demonstrate orthogonality between UV-B, blue, and red/far-red light-responsive gene switches in a single mammalian cell culture. We expect this approach to enable the spatiotemporal control of gene networks and to expand the applications of optogenetics in synthetic biology.
光遗传学基因开关能够以前所未有的时空分辨率控制基因表达。最近，已开发出由UV-B、蓝光或红光激活的光响应转基因表达系统。这些系统自身表现良好，但由于光感受器的吸收光谱重叠，它们整合到遗传网络中受到了阻碍。我们发现UV-B和蓝光控制的基因表达之间缺乏正交性是瓶颈所在，并采用了基于模型的方法，该方法确定需要一种对低强度光不敏感的蓝光响应基因开关。基于这一预测，我们开发了一种蓝光响应且快速可逆的表达系统。最后，我们利用该表达系统在单一哺乳动物细胞培养中证明了UV-B、蓝光和红/远红光响应基因开关之间的正交性。我们期望这种方法能够实现基因网络的时空控制，并扩展光遗传学在合成生物学中的应用。
