The aim of this project is to identify active substances that specifically prevent the packaging of the SARS-CoV-2 genome into the viral particle and thus inhibiting its replication. Packaging of the viral genome into the virus is an essential process in the viral life cycle whereby the nucleocapsid protein (N) specifically binds to the RNA genome and organizes its packaging in the virus. In particular, SARS N undergoes liquid-liquid phase separation (LLPS) with RNA, which contributes to viral transcription, replication and ultimately virion assembly.
In this project, a structure- based and cellular screening system will be established that can identify molecules that interact with N. Moreover, the functional effects of these interactors on RNA-binding and packaging will be analysed. Ultimately, to determine effective therapeutics, their impact on viral replication and infectivity will be directly assessed in cellular systems. For this we already successfully established a stable cell line that can be induced by doxocycline to express SARS-CoV-2 N protein. This cell line is the basis for the establishment of a cell-based screening assay using phase separation as a readout.
AG Längst, Fraunhoferinstitut
University of Regensburg, Institute for Medical Microbiology and Hygiene