SARS-CoV-2 RNA Polymerase (RdRp) Assays for Drug Discovery

The COVID-19 pandemic is caused by infection of SARS-CoV-2 or severe acute respiratory syndrome coronavirus 2, a positive-sense single-stranded RNA virus. The RNA dependent RNA polymerase (RdRp) is a key drug target for this deadly virus. The high throughput SARS-CoV-2 RdRp assay is based on detection of the RNA molecules synthesized by the RNA polymerase in the presence of an RNA template and NTPs. The assay can be performed in a 384-well or 96-well plate format for tests of the enzyme activities of the virus RNA polymerase (SARS-CoV-2 RdRp) and high throughput screening of inhibitors.   

The gel-based assay is also available for visualization of the RNA product synthesized by RdRp and inhibition by inhibitors.

High throughput assay kits:

The SARS-CoV-2 RNA polymerase assay kit-100 (Catalog No.S2RPA100K) includes 300 µl of 10 x Buffer, 50 µl of 50 x RNA template, 60 µl of 50 x  mM NTPs (ATP, UTP, GTP, CTP) and 700 µl of 10 x fluorescence dye. It is for 100 assays of SARS-CoV-2 RNA polymerase in a 384-well plate format or 50 assays in a 96-well plate format. The assay kit includes all reagents except the enzyme. 

The SARS-CoV-2 RNA polymerase assay kit plus-20 (Catalog No.S2RPA020KE) includes 75 µl of 10 x Buffer,  10 µl of 50 x RNA template, 12 µl of 50 x NTPs (ATP, UTP, GTP, CTP), 150 µl of 10 x fluorescence dye and  10 µl of 50 x SARS-CoV-2 RdRp.  It is for 20 assays of SARS-CoV-2 RNA polymerase in a 384-well plate format or 10 assays in a 96-well plate format.  

The SARS-CoV-2 RNA polymerase assay kit plus-100 (Catalog No.S2RPA100KE) includes 300 µl of 10 x Buffer, 50 µl of 50 x RNA template, 60 µl of 50 x NTPs (ATP, UTP, GTP, CTP), 700 µl of 10 x fluorescence dye and  50 µl of 50 x SARS-CoV-2 RdRp.  It is for 100 assays of SARS-CoV-2 RNA polymerase in a 384-well plate format or 50 assays in a 96-well plate format.

Gel-based assay kit:

The Gel-based SARS-CoV-2 RdRp assay kit (Catalog No. GS2RPA020KE) includes all the reagents for 20 samples in gel-based assays of SARS-CoV-2 RdRp activity.  It includes 75 µl of 10 x Buffer, 10 µl of 50 x RNA template, 12 µl of 50 x NTPs (ATP, UTP, GTP, CTP), 10 µl of 50 x SARS-CoV-2 RdRp and 130 µl of 5 x Gel loading buffer.

Functional SARS-CoV-2 RdRp enzyme

The SARS-CoV-2 RdRp complex – for 100 assays (Catalog No. RDRP-100S2) includes 50 µl of 50 x RNA-dependent RNA Polymerase composed of the complex of the nsp12, nsp7 and nsp8 proteins from SARS-CoV-2 virus. It is for 100 assays in a 384-well plate assay format (25 µl reaction) or 50 assays in a 96-well plate format (50 µl reaction).

Publications:

Pacl H. T et al, Water-soluble tocopherol derivatives inhibit SARS-CoV-2 RNA-dependent RNA polymerase. bioRxiv, Cold Spring Harbor Laboratory, July 27, 2021.

Goc A. et al, Simultaneous Inhibition of SARS-CoV-2 Infectivity by a Specific Combination of Plant-derived Compounds. European Journal of Biology and Biotechnology, Vol 2,  Issue 5, p25 (2021). 

Goc A. et al, Inhibitory effects of specific combination of natural compounds against SARS-CoV-2 and its Alpha, Beta, Gamma, Delta, Kappa, and Mu variants. European Journal of Microbiology and Immunology, 21 Jan 2022.