Photoactivated RNA Binding in a Blue-Light Receptor Enables Optoribogenetics

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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.