Photoactivated RNA Binding in a Blue-Light Receptor Enables Optoribogenetics

蓝光受体中的光激活 RNA 结合使光遗传学成为可能

• 批准号: 315180630
• 负责人: Professor Dr. Günter Mayer
• 金额: --
• 依托单位: Arbeitsgruppe Biochemie I
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2016
• 资助国家: 德国
• 起止时间: 2015-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/315180630?language=en
• 关键词: Photoactivated; RNA; Binding; Blue; Light

Sensory photoreceptors underlie various light-dependent organismal adaptations in nature. In optogenetics, photoreceptors double as genetically encodable actuators for the control of cellular and organismal physiology with reversibility, spatiotemporal resolution and minimal invasiveness. Naturally occurring photoreceptors and a plethora of engineered representatives now enable ver-satile optogenetic intervention into manifold cellular processes. That notwithstanding, there has been a dearth of optogenetic actuators that would directly act on RNA. We have recently closed this gap with the discovery of the light-oxygen-voltage (LOV) receptor PAL from the actinobacte-rium Nakamurella multipartita. PAL sequence-specifically binds short RNA aptamers with affinity that is enhanced by around 100-fold under blue light and thereby enables hitherto inaccessible optoribogenetic applications. The current, collaborative proposal aims at obtaining a molecular understanding of light regulation and RNA binding in PAL and two recently discovered homologs. Pertinent insight informs the engineering of PAL derivatives with altered and enhanced traits, par-ticularly RNA sequence specificity and response to light. We will harness the light-regulated PAL:aptamer interaction for the spatiotemporal control of micro RNA splicing and expression reg-ulation of endogenous genes, both in cell culture and in vivo. The planned research paves the way towards novel light-regulated actuators suitable for the precise optoribogenetic interrogation of the myriad roles RNA fulfills in nature.

感觉光感受器是自然界中各种光依赖性生物适应的基础。在光遗传学中，光感受器兼作遗传编码的致动器，用于控制具有可逆性、时空分辨率和最小侵入性的细胞和有机体生理学。天然存在的光感受器和过多的工程化代表现在使多功能光遗传学干预成为多种细胞过程。尽管如此，仍然缺乏直接作用于RNA的光遗传致动器。最近，我们从放线菌Nakamurella multipartita中发现了光氧电压（LOV）受体PAL，从而填补了这一空白。PAL序列特异性结合短RNA适体，其亲和力在蓝光下增强约100倍，从而实现迄今为止无法实现的光生应用。目前，合作的建议旨在获得光调节和PAL和两个最近发现的同系物的RNA结合的分子理解。相关的见解告知PAL衍生物的工程改造，具有改变和增强的性状，特别是RNA序列特异性和对光的反应。我们将利用光调控PAL：适体相互作用的时空控制的微小RNA剪接和内源性基因的表达调控，在细胞培养和体内。这项计划中的研究为新型光调控致动器铺平了道路，这些致动器适用于对RNA在自然界中所扮演的无数角色进行精确的optoribogenetic询问。