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Citation: Yuan Zhang, Chunjie Li, Xianliang Ke, Dan Luo, Yan Liu, Quanjiao Chen, Hanzhong Wang, Xiaohui Song, Zhenhua Zheng. Development of a biosensor assessing SARS-CoV-2 main protease proteolytic activity in living cells for antiviral drugs screening [J].VIROLOGICA SINICA, 2022, 37(3) : 459-461.  http://dx.doi.org/10.1016/j.virs.2022.04.002

Development of a biosensor assessing SARS-CoV-2 main protease proteolytic activity in living cells for antiviral drugs screening

  • Highlights:
    The biosensor reported in our study can monitor SARS-CoV-2 Mpro activity in living cells instead of in vitro solutions.
    The biosensor reported in our study is sensitive and easy to operate.
    It is suitable for high-throughput screening.
    It has the potential to be used in small animal models.

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  • 10.1016j.virs.2022.04.002-ESM.docx
    1. Bartok, E., Bauernfeind, F., Khaminets, M.G., Jakobs, C., Monks, B., Fitzgerald, K.A., Latz, E., Hornung, V., 2013. iGLuc:a luciferase-based inflammasome and protease activity reporter. Nat. Methods 10, 147-154.

    2. Bowlby, M.R., Case, J.F., 1991. Flash kinetics and spatial patterns of bioluminescence in the copepod gaussia-princeps. Mar. Biol. 110, 329-336.

    3. Froggatt, H.M., Heaton, B.E., Heaton, N.S., 2020. Development of a fluorescence-based, high-throughput SARS-CoV-2 3CL(pro) reporter assay. J. Virol. 94, e01265, 20.

    4. Jin, Z., Du, X., Xu, Y., Deng, Y., Liu, M., Zhao, Y., Zhang, B., Li, X., Zhang, L., Peng, C., Duan, Y., Yu, J., Wang, L., Yang, K., Liu, F., Jiang, R., Yang, X., You, T., Liu, X., Yang, X., Bai, F., Liu, H., Liu, X., Guddat, L.W., Xu, W., Xiao, G., Qin, C., Shi, Z., Jiang, H., Rao, Z., Yang, H., 2020. Structure of M(pro) from SARS-CoV-2 and discovery of its inhibitors. Nature 582, 289-293.

    5. Pillaiyar, T., Manickam, M., Namasivayam, V., Hayashi, Y., Jung, S.H., 2016. An overview of severe acute respiratory syndrome-coronavirus (SARS-CoV) 3CL protease inhibitors:peptidomimetics and small molecule chemotherapy. J. Med. Chem. 59, 6595-6628.

    6. Rawson, J.M.O., Duchon, A., Nikolaitchik, O.A., Pathak, V.K., Hu, W.S., 2021. Development of a cell-based luciferase complementation assay for identification of SARS-CoV-2 3CL(pro) inhibitors. Viruses 13, 173.

    7. Wu, F., Zhao, S., Yu, B., Chen, Y.M., Wang, W., Song, Z.G., Hu, Y., Tao, Z.W., Tian, J.H., Pei, Y.Y., Yuan, M.L., Zhang, Y.L., Dai, F.H., Liu, Y., Wang, Q.M., Zheng, J.J., Xu, L., Holmes, E.C., Zhang, Y.Z., 2020. A new coronavirus associated with human respiratory disease in China. Nature 579, 265-269.

    8. Zhou, P., Yang, X.L., Wang, X.G., Hu, B., Zhang, L., Zhang, W., Si, H.R., Zhu, Y., Li, B., Huang, C.L., Chen, H.D., Chen, J., Luo, Y., Guo, H., Jiang, R.D., Liu, M.Q., Chen, Y., Shen, X.R., Wang, X., Zheng, X.S., Zhao, K., Chen, Q.J., Deng, F., Liu, L.L., Yan, B., Zhan, F.X., Wang, Y.Y., Xiao, G.F., Shi, Z.L., 2020. A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature 579, 270-273.

    9. Zhu, W., Xu, M., Chen, C.Z., Guo, H., Shen, M., Hu, X., Shinn, P., Klumpp-Thomas, C., Michael, S.G., Zheng, W., 2020. Identification of SARS-CoV-2 3CL protease inhibitors by a quantitative high-throughput screening. ACS Pharmacol Transl Sci 3, 1008-1016.

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    Development of a biosensor assessing SARS-CoV-2 main protease proteolytic activity in living cells for antiviral drugs screening

      Corresponding author: Xiaohui Song, songxiaohui@zgwhfe.com
      Corresponding author: Zhenhua Zheng, zhengzh@wh.iov.cn
    • a Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China;
    • b Department of Gastroenterology, Wuhan Children's Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, 430015, China;
    • c Department of Obstetrics, Wuhan Children's Hospital, Wuhan Maternal and Child Healthcare Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, 430015, China;
    • d University of the Chinese Academy of Sciences, Beijing, 100049, China

    Abstract: Highlights:
    The biosensor reported in our study can monitor SARS-CoV-2 Mpro activity in living cells instead of in vitro solutions.
    The biosensor reported in our study is sensitive and easy to operate.
    It is suitable for high-throughput screening.
    It has the potential to be used in small animal models.

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