Periodic Reporting for period 4 - RNArepair (Site-directed RNA Editing to Manipulate RNA and Protein Function)
Berichtszeitraum: 2020-02-01 bis 2021-01-31
However, to make such tools efficient and precise is a major challenge that requires to combine expertise from chemistry and life sciences. Coming from a chemistry background, we have established an entirely novel access to assemble an editing tool inside the cell. Within this project we aim to better understand our RNa targeting strategy by comprehensively characterizing all its properties. Furthermore, we aim to develop several alternative methods to deliver the tool into relevant target tissues. Finally, we wish to implement the tool in vivo. The project aims to make the basic research community and biotech companies familiar with the novel RNA targeting approach and to foster the development of site-directed RNA base editing as a platform for therapy and advanced RNA-targeting strategies.
We produced the various viral vehicles (lenti and adeno virus) to become more flexible with the delivery of the tool. This allowed us to apply the tool in fragile and difficult to transfect cells like primary cells. We also achieved to stabilize the guideRNA for naked and receptor-mediated uptake.
Finally, we implemented two photocontrol strategies and tested them successfully in cell culture and in vivo in an annelid (Platynereis dumerilii). Furthermore, we also implemented RNA editing for the inclusion N- and C-terminal protein localization signals and achieved photocontrol over protein localization. We were able to perturb signaling cues by site-directed RNA editing.
The results of the action are partially published, e.g. in the internationally highly visible journal Nature Methods. The results were presented on numerous internal conferences and workshops, including the Annual Meeting of the Oligonucleotide Therapeutics Society in 2019, the 2020 Nature Conference on RNA "from bench to bedside", and the joint ERC-EIC workshop on Gene and Cell Therapy in 2021. The work has been highlighted by several features, including a highlight in Nature in 2020. RNA base editing became a recent focus of biotech industry with several newly formed companies working at the clinical translation of the technology, as featured by Chemical&Engineering News in 2019.