This study by Ernawati Arifin Giri-Rachman, Ph.D., Azzania Fibriani, M.Si., Ph.D., and their colleagues aimed to develop a novel SARS-CoV-2 main protease (Mpro) dimer-based screening system (DBSS) utilizing synthetic biology in Escherichia coli BL21(DE3). We linked the SARS-CoV-2 Mpro with the DNA-binding domain of AraC regulatory protein, which regulates the reporter gene expression. Protein modeling and molecular docking showed that saquinavir could bind to AraC-Mpro both in its monomer and dimer forms. The constructed DBSS assay indicated the screening system could detect saquinavir inhibitory activity at a concentration range of 4–10 μg/mL compared to the untreated control (P ≤ 0.05). The Vero E6 cell assay validated the DBSS result that saquinavir at 4–10 μg/mL exhibited antiviral activity against SARS-CoV-2. Our DBSS could be used for preliminary screening of numerous drug candidates that possess a dimerization inhibitor activity of SARS-CoV-2 Mpro and also minimize the use of a high biosafety level laboratory.

Article Citation:
Giri-Rachman, E. A., Effendy, V.V., Azmi, M.H.S., Yamahoki, N., Stephanie, R., Agustiyanti, D.F., Wisnuwardhani, P.H., Angelina, M., Rubiyana, Y., Aditama, R., Ningrum, R.A., Wardiana, A., Fibriani, A. (2024). The SARS-CoV-2 Mpro Dimer-Based Screening System: A Synthetic Biology Tool for Identifying Compounds with Dimerization Inhibitory Potential. ACS Synth. Biol. 13(2): 509–520. https://doi.org/10.1021/acssynbio.3c00446