Design of modular autoproteolytic gene switches responsive to anti-coronavirus drug candidates

被引:22
作者
Franko, Nik [1 ]
Teixeira, Ana Palma [1 ]
Xue, Shuai [1 ]
Charpin-El Hamri, Ghislaine [2 ]
Fussenegger, Martin [1 ,3 ]
机构
[1] Swiss Fed Inst Technol, Dept Biosyst Sci & Engn, Mattenstr 26, CH-4058 Basel, Switzerland
[2] Univ Claude Bernard Lyon 1, Inst Univ Technol, Dept Genie Biol, F-69622 Villeurbanne, France
[3] Univ Basel, Fac Life Sci, Basel, Switzerland
基金
瑞士国家科学基金会;
关键词
HIV PROTEASE INHIBITORS; PAPAIN-LIKE; IDENTIFICATION;
D O I
10.1038/s41467-021-27072-3
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
SARS-CoV-2 proteases are key targets for anti-viral drug development. Here the authors present modular tunable autoproteolytic gene switches for virus free cell culture screening of inhibitors. The main (Mpro) and papain-like (PLpro) proteases encoded by SARS-CoV-2 are essential to process viral polyproteins into functional units, thus representing key targets for anti-viral drug development. There is a need for an efficient inhibitor screening system that can identify drug candidates in a cellular context. Here we describe modular, tunable autoproteolytic gene switches (TAGS) relying on synthetic transcription factors that self-inactivate, unless in the presence of coronavirus protease inhibitors, consequently activating transgene expression. TAGS rapidly report the impact of drug candidates on Mpro and PLpro activities with a high signal-to-noise response and a sensitivity matching concentration ranges inhibiting viral replication. The modularity of the TAGS enabled the study of other Coronaviridae proteases, characterization of mutations and multiplexing of gene switches in human cells. Mice implanted with Mpro or PLpro TAGS-engineered cells enabled analysis of the activity and bioavailability of protease inhibitors in vivo in a virus-free setting.
引用
收藏
页数:12
相关论文
共 46 条
[1]  
Ausländer D, 2018, NAT METHODS, V15, P57, DOI [10.1038/nmeth.4505, 10.1038/NMETH.4505]
[2]   Efficacy and Safety of the mRNA-1273 SARS-CoV-2 Vaccine [J].
Baden, Lindsey R. ;
El Sahly, Hana M. ;
Essink, Brandon ;
Kotloff, Karen ;
Frey, Sharon ;
Novak, Rick ;
Diemert, David ;
Spector, Stephen A. ;
Rouphael, Nadine ;
Creech, C. Buddy ;
McGettigan, John ;
Khetan, Shishir ;
Segall, Nathan ;
Solis, Joel ;
Brosz, Adam ;
Fierro, Carlos ;
Schwartz, Howard ;
Neuzil, Kathleen ;
Corey, Larry ;
Gilbert, Peter ;
Janes, Holly ;
Follmann, Dean ;
Marovich, Mary ;
Mascola, John ;
Polakowski, Laura ;
Ledgerwood, Julie ;
Graham, Barney S. ;
Bennett, Hamilton ;
Pajon, Rolando ;
Knightly, Conor ;
Leav, Brett ;
Deng, Weiping ;
Zhou, Honghong ;
Han, Shu ;
Ivarsson, Melanie ;
Miller, Jacqueline ;
Zaks, Tal .
NEW ENGLAND JOURNAL OF MEDICINE, 2021, 384 (05) :403-416
[3]   The genetic design of signaling cascades to record receptor activation [J].
Barnea, Gilad ;
Strapps, Walter ;
Herrada, Gilles ;
Berman, Yemiliya ;
Ong, Jane ;
Kloss, Brian ;
Axel, Richard ;
Lee, Kevin J. .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2008, 105 (01) :64-69
[4]   High-throughput screening identifies inhibitors of the SARS coronavirus main proteinase [J].
Blanchard, JE ;
Elowe, NH ;
Huitema, C ;
Fortin, PD ;
Cechetto, JD ;
Eltis, LD ;
Brown, ED .
CHEMISTRY & BIOLOGY, 2004, 11 (10) :1445-1453
[5]   A conformational switch high-throughput screening assay and allosteric inhibition of the flavivirus NS2B-NS3 protease [J].
Brecher, Matthew ;
Li, Zhong ;
Liu, Binbin ;
Zhang, Jing ;
Koetzner, Cheri A. ;
Alifarag, Adham ;
Jones, Susan A. ;
Lin, Qishan ;
Kramer, Laura D. ;
Li, Hongmin .
PLOS PATHOGENS, 2017, 13 (05)
[6]   Engineering protein-protein devices for multilayered regulation of mRNA translation using orthogonal proteases in mammalian cells [J].
Cella, Federica ;
Wroblewska, Liliana ;
Weiss, Ron ;
Siciliano, Vera .
NATURE COMMUNICATIONS, 2018, 9
[7]   Profiling of Substrate Specificity of SARS-CoV 3CLpro [J].
Chuck, Chi-Pang ;
Chong, Lin-Tat ;
Chen, Chao ;
Chow, Hak-Fun ;
Wan, David Chi-Cheong ;
Wong, Kam-Bo .
PLOS ONE, 2010, 5 (10)
[8]  
Chung HK, 2015, NAT CHEM BIOL, V11, P713, DOI [10.1038/NCHEMBIO.1869, 10.1038/nchembio.1869]
[9]  
ClinicalTrials.gov, 2020, SINGL ASC DOS STUD I
[10]  
Cohen J., 2020, SCIENCE, DOI [10.1126/science.abf4549, DOI 10.1126/SCIENCE.ABF4549]