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OptoRibo Report Summary

Project ID: 615381
Funded under: FP7-IDEAS-ERC
Country: Germany

Mid-Term Report Summary - OPTORIBO (Optogenetic control of cellular behaviour by allosteric ribonucleic acid assemblies)

The overall goal of OptoRibo is to generate novel RNA assemblies in which individual RNA domains are under optogenetic control. We seek to obtain and explore modularly assembled RNA devices with novel functions for the analysis of endogenous, intracellular biomolecules to investigate processes at the level of cells, tissue and entire organisms with high spatiotemporal resolution.
To achieve this, the OptoRibo project is divided in three major objectives. The first objective covers the generation of aptamers that recognise photoreceptor proteins (PRP) in a light-dependent manner. It comprises the characterisation of obtained aptamers in regard of biophysical properties, e.g. the determination of individual dissociation constants, structure-activity relationship and performance depending on the irradiation status of the respective PRP.
The second objective comprises the generation of RNA devices and allosteric RNA assemblies that are built from the PRP-specific aptamers and a second RNA domain. Notably, these two RNA domains will be conjoined in an allosteric manner, hence the binding status of the PRP-binding domain will dictate the property and performance of the joint second RNA domain. In the third objective, we apply these allosteric RNA assemblies and devices to cellular systems and living model organisms, e.g. flies and mice.
In summary, we have made substantial progress in the OptoRibo project and met all of the anticipated aims. Most importantly, we identified a very short aptamer that binds to a PRP in a light-dependent manner. This aptamer is a valuable starting point for cellular and in vivo studies. Besides the proposed strategies, i.e. regulation of siRNA and aptamer function we included further applications of the PRP-aptamer in our endeavours. This will allow to broadening the approach and opens further opportunities, e.g. optogenetic RNA tagging and tracking and photochemical genetic engineering, both of which are unprecedented.

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