RBD - RNA binding inhibitor screen - project

RBD is one of two RNA binding domains of NP - Inhibition of RNA binding should affect the viral life cycle

Ongoing work and plans

PharmAI screened the RBD of Nucleocapsid - RBD inhibitors 1st round - PharmAI

Summary of results and identified inhibitors can be found here: List of RNA-binding inhibitors found

Key Results

we identified RBD binders in vitro; if they really inhibit RNA binding is not so clear yet

MST is not the method of choice for screening - RBD - MST inhibitor screen

nanoDSF was good for screening - RBD - nanoDSF screens

some of these work in vivo: List of RNA-binding inhibitors found](https://app.thebrain.com/brain/7b136aa7-aacb-44e7-9d7e-b6204ba2fc9f/e6b2f5ce-6ebd-4a9a-87a6-0225d6bf69f7)

Next Steps

NMR to show binding (Remco - did not work)

On/OFF rates (WAVE -2bind - did not work)

Inhibition of LLPS by these inhibitors - (in vitro assay - Niklas ongoing; in vivo Fraunhofer ongoing)

measuring RBD RNA binding affinity in the presence of inhbitor by nanoDSF - see: PMID: 30414312 (no effect - Niklas)

Effect on RNA binding - EMSA

The inhibitors could inhibit the chaperoning activity - see: NP - RBD chaperone in RNA folding. additional questions. is the full length protein required for chaperoning? what about mixing rbd+ctd?