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Various human outbreaks occurred in the 1980s, resulting from a cessation of smallpox vaccination. Recently, MPXV cases have reemerged in non\u2010endemic nations, and the 2022 outbreak has been declared a public health emergency. Treatment optionsare limited, and many countries lack the infrastructure to provide symptomatic treatments. The development of cost\u2010effective antivirals could ease severe health outcomes. G\u2010quadruplexes have been a target of interest in treating viral infections with different chemicals. In the present work, a genomic\u2010scale mapping of different MPXV isolates highlighted two conserved putative quadruplex\u2010forming sequences MPXV\u2010exclusive in 590 isolates. Subsequently, we assessed the G\u2010quadruplex formation using circular dichroism\u00a0spectroscopy and solution small\u2010angle X\u2010ray scattering. Furthermore, biochemical assays indicated the ability of MPXV quadruplexes to be recognized by two specific G4\u2010binding partners\u2014Thioflavin T and DHX36. Additionally, our work also suggests that a quadruplex binding small\u2010molecule with previously reported antiviral activity, TMPyP4, interacts with MPXV G\u2010quadruplexes with nanomolar affinity in the presence and absence of DHX36. Finally, cell biology experiments suggests that TMPyP4 treatment substantially reduced gene expression of MPXV proteins. In summary, our work provides insights into the G\u2010quadruplexes from the MPXV genome that can be further exploited to develop therapeutics.<\/jats:p>","DOI":"10.1002\/jmv.28783","type":"journal-article","created":{"date-parts":[[2023,5,10]],"date-time":"2023-05-10T13:48:50Z","timestamp":1683726530000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Mapping and characterization of G\u2010quadruplexes in monkeypox genomes"],"prefix":"10.1002","volume":"95","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4761-8361","authenticated-orcid":false,"given":"Higor Sette","family":"Pereira","sequence":"first","affiliation":[{"name":"Department of Chemistry and Biochemistry, Alberta RNA Research and Training Institute University of Lethbridge Lethbridge Alberta Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2834-9063","authenticated-orcid":false,"given":"Darren L.","family":"Gemmill","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry, Alberta RNA Research and Training Institute University of Lethbridge Lethbridge Alberta Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0891-3122","authenticated-orcid":false,"given":"M. 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