Circularly permuted LOV2 as a modular photoswitch for optogenetic engineering
Circularly permuted LOV2 as a modular photoswitch for optogenetic engineering
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DOI：
10.1038/s41589-021-00792-9
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发表时间：
2021-05-06
影响因子：
14.8
通讯作者：
Zhou, Yubin
中科院分区：
文献类型：
--
作者：
He, Lian;Tan, Peng;Zhou, Yubin
基于植物的光传感器，例如燕麦趋光素 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蔽表面选择有限及其偏好容纳 C 端 J α 螺旋下游效应器结构域的阻碍。在这里，我们设计了一组具有额外笼养功能的 LOV2 圆形排列（cpLOV2），从而扩展了基因编码光开关的功能，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、引发细胞自杀并生成可光激活的嵌合抗原受体 T 细胞以诱导肿瘤细胞杀伤。我们的方法广泛适用于工程其他光感受器，以满足针对生物医学和生物技术应用定制的光遗传学工具日益增长的需求。一组LOV2圆形排列（cpLOV2）已被设计用于扩展现有的光遗传学工具箱。 cpLOV2 能够设计 ON 和 OFF 开关来控制细胞信号传导、基因表达、细胞命运和癌症免疫治疗。
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 J alpha 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.A set of LOV2 circular permutants (cpLOV2) have been engineered to expand the existing optogenetic toolbox. cpLOV2 enables the design of both ON- and OFF-switches to control cell signaling, gene expression, cell fate and cancer immunotherapy.