Plant-based photosensors, such as the light-oxygen-voltage sensing domain 2 (LOV2) from oat phototropin 1, can be modularly wired into cell signaling networks to remotely control protein activity and physiological processes. However, the applicability of LOV2 is hampered by the limited choice of available caging surfaces and its preference to accommodate the effector domains downstream of the C-terminal Jα helix. Here, we engineered a set of LOV2 circular permutants (cpLOV2) with additional caging capabilities, thereby expanding the repertoire of genetically encoded photoswitches to accelerate the design of optogenetic devices. We demonstrate the use of cpLOV2-based optogenetic tools to reversibly gate ion channels, antagonize CRISPR–Cas9-mediated genome engineering, control protein subcellular localization, reprogram transcriptional outputs, elicit cell suicide and generate photoactivatable chimeric antigen receptor T cells for inducible tumor cell killing. Our approach is widely applicable for engineering other photoreceptors to meet the growing need of optogenetic tools tailored for biomedical and biotechnological applications.

基于植物的光传感器，例如来自燕麦 phototropin 1 的光氧电压传感域 2 (LOV2)，可以模块化连接到细胞信号网络中，以远程控制蛋白质活性和生理过程。然而，LOV2 的适用性受到可用笼罩表面的有限选择及其偏好容纳 C 末端 Jα 螺旋下游的效应域的阻碍。在这里，我们设计了一组 LOV2 循环置换 (cpLOV2) 具有额外的笼子功能，从而扩展了基因编码的光开关的全部内容，以加速光遗传学设备的设计。我们展示了使用基于 cpLOV2 的光遗传学工具来可逆地门控离子通道，拮抗 CRISPR-Cas9 介导的基因组工程，控制蛋白质亚细胞定位，重新编程转录输出，引发细胞自杀并产生可光激活的嵌合抗原受体 T 细胞，用于诱导肿瘤细胞杀伤。我们的方法广泛适用于对其他光感受器进行工程设计，以满足为生物医学和生物技术应用量身定制的光遗传学工具日益增长的需求。