{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T12:31:59Z","timestamp":1792672319366,"version":"4.5.0"},"update-to":[{"DOI":"10.1371\/journal.ppat.1013764","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T00:00:00Z","timestamp":1773014400000}}],"reference-count":27,"publisher":"Public Library of Science (PLoS)","issue":"3","license":[{"start":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T00:00:00Z","timestamp":1772496000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["223608\/Z\/21\/Z, 217105\/Z\/19\/Z"],"award-info":[{"award-number":["223608\/Z\/21\/Z, 217105\/Z\/19\/Z"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plospathogens.org"],"crossmark-restriction":false},"short-container-title":["PLoS Pathog"],"abstract":"<jats:p>\n                    Acoziborole is a safe, single\u00a0dose, oral therapy, for treatment of both early and late-stage sleeping sickness, a deadly disease caused by African trypanosomes. Other benzoxaboroles show efficacy against other trypanosomatids, apicomplexans, fungi, bacteria, and viruses. Acoziborole targets the trypanosome pre-mRNA processing endonuclease, cleavage and polyadenylation specificity factor 3 (CPSF3), and triggers CPSF3 degradation, but it remains unclear whether additional mechanisms contribute to efficacy. We used oligo targeting for site saturation mutagenesis of the native\n                    <jats:italic>CPSF3<\/jats:italic>\n                    gene. Among &gt;1,500 edits around the putative drug binding site, only Asn\n                    <jats:sup>232<\/jats:sup>\n                    His edits conferred moderate resistance to acoziborole. Using a combinatorial oligo targeting method we edited multiple sites simultaneously, including sites that differ in human CPSF3, and found that an Asn\n                    <jats:sup>232<\/jats:sup>\n                    His, Tyr\n                    <jats:sup>383<\/jats:sup>\n                    Phe, Asn\n                    <jats:sup>448<\/jats:sup>\n                    Gln triple-mutant strain was\u2009&gt;\u200940-fold resistant to acoziborole. We used gene tagging to show that all three edits were on the same allele, and to show that triple-mutant CPSF3 was highly resistant to rapid acoziborole and proteasome-dependent degradation. Computational modelling revealed how the combinatorial mutations can disrupt acoziborole \u2013 CPSF3 interactions by introducing steric clash and by disrupting hydrophobic and water-mediated interactions. We conclude that acoziborole safety and efficacy can be explained by selective affinity for, and rapid turnover of, trypanosome CPSF3.\n                  <\/jats:p>","DOI":"10.1371\/journal.ppat.1013764","type":"journal-article","created":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T18:32:53Z","timestamp":1772562773000},"page":"e1013764","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":3,"title":["Acoziborole resistance associated mutations in Trypanosoma brucei 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