2. Academic Validation
  3. Development of a replication-defective mpox virus platform for fundamental and therapeutic research

Development of a replication-defective mpox virus platform for fundamental and therapeutic research

  • Nat Commun. 2025 Dec 15. doi: 10.1038/s41467-025-67487-w.
Jiannan Chen # 1 2,  Liyuan Hu # 1,  Riccardo Vernuccio # 3,  Ning Shi # 4,  Jiaxin Tian 5,  Yuyi Zhang 1,  Sicheng Tian 1,  Xinyu Cao 5,  Zhuo Ha 5,  Jiahan Lu 6,  Leandro Battini 3,  Bertrand Raynal 7,  Ahmed Haouz 8,  Jing Xue 6,  Qiliang Cai 1,  Yiqi Zhao 9,  Yongxu Lu 10,  Geoffrey L Smith 9 10 11,  Youhua Xie 12,  Huijun Lu 13,  Pablo Guardado-Calvo 14,  Ping Zhang 15,  Rong Zhang 16
Affiliations

Affiliations

  • 1 Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), Shanghai Institute of Infectious Disease and Biosecurity, Shanghai Frontiers Science Center of Pathogenic Microorganisms and Infection, School of Basic Medical Sciences, Fudan University, Shanghai, China.
  • 2 Department of Immunology and Microbiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.
  • 3 G5 Structural Biology of Infectious Diseases, Institut Pasteur, Université Paris Cité, Paris, France.
  • 4 State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, Institute of Zoonosis, and College of Veterinary Medicine, Jilin University, Changchun, Jilin Province, China.
  • 5 Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun, Jilin Province, China.
  • 6 State Key Laboratory of Respiratory Health and Multimorbidity, NHC Key Laboratory of Human Disease Comparative Medicine, Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
  • 7 Plate-Forme de Biophysique Moleculaire-C2RT, Institut Pasteur, CNRS UMR 3528, Université Paris Cité, Paris, France.
  • 8 Crystallography Platform-C2RT, UMR 3528, Institut Pasteur, CNRS, Université de Paris, Paris, France.
  • 9 Sir William Dunn School of Pathology, University of Oxford, Oxford, UK.
  • 10 Vaccine Research Centre, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China.
  • 11 Chinese Academy of Medical Sciences-Oxford Institute, University of Oxford, Oxford, UK.
  • 12 Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), Shanghai Institute of Infectious Disease and Biosecurity, Shanghai Frontiers Science Center of Pathogenic Microorganisms and Infection, School of Basic Medical Sciences, Fudan University, Shanghai, China. [email protected].
  • 13 Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun, Jilin Province, China. [email protected].
  • 14 G5 Structural Biology of Infectious Diseases, Institut Pasteur, Université Paris Cité, Paris, France. [email protected].
  • 15 Department of Immunology and Microbiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China. [email protected].
  • 16 Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), Shanghai Institute of Infectious Disease and Biosecurity, Shanghai Frontiers Science Center of Pathogenic Microorganisms and Infection, School of Basic Medical Sciences, Fudan University, Shanghai, China. [email protected].
  • # Contributed equally.
PMID: 41397984 DOI: 10.1038/s41467-025-67487-w
Abstract

The recent global outbreaks of mpox highlight the urgent need for both fundamental research and Antiviral development. However, studying the mpox virus (MPXV), with its large and complex genome, remains challenging due to the requirement for high-containment facilities. Here, we describe a strategy for de novo assembly of MPXV clade IIb genomes in Bacterial artificial chromosomes using transformation-associated recombination cloning. Leveraging CRISPR-Cas9 and Lambda Red recombination, we engineer replication-defective MPXV particles with dual deletions of OPG96 (M2R) and OPG158 (A32.5 L)-genes essential for virion assembly, that are capable of recapitulating key stages of the viral life cycle. We apply this system to screen a compound library and identify G243-1720, a potent anti-poxvirus inhibitor with broad activity in vitro and in vivo. G243-1720 blocks the formation of extracellular enveloped virions and cell-cell spread. Resistance mutation selection, crystallographic analysis, analytical ultracentrifugation, and mass photometry reveal that, despite its distinct chemical structure, G243-1720 shares a mode of action with tecovirimat, both functioning by affecting dimerization of protein OPG57 (F13). Our findings underscore the potential of G243-1720 as a promising broad-spectrum anti-poxvirus lead compound and demonstrate the utility of replication-defective MPXV particles as a reliable platform for viral biology studies and Antiviral development.

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