{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T12:57:14Z","timestamp":1753880234588,"version":"3.41.2"},"reference-count":42,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2025,4,2]],"date-time":"2025-04-02T00:00:00Z","timestamp":1743552000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,4,2]],"date-time":"2025-04-02T00:00:00Z","timestamp":1743552000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"crossref","award":["JP23fk0108680"],"award-info":[{"award-number":["JP23fk0108680"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"crossref","award":["JP23wm0225036"],"award-info":[{"award-number":["JP23wm0225036"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001691","name":"MEXT | Japan Society for the Promotion of Science","doi-asserted-by":"crossref","award":["18K07095"],"award-info":[{"award-number":["18K07095"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2025,4,2]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>\n              Malaria parasites acquire drug resistance through genetic changes, the mechanisms of which remain incompletely understood. Understanding the mechanisms of drug resistance is crucial for the development of effective treatments against malaria, and for this purpose, new genetic tools are needed. In a previous study, as a forward genetic tool, we developed the rodent malaria parasite\n              <jats:italic>Plasmodium berghei<\/jats:italic>\n              mutator (PbMut) line, which has a greatly increased rate of mutation accumulation and from which we isolated a mutant with reduced susceptibility to piperaquine (PPQ). We identified a mutation in the\n              <jats:italic>chloroquine resistance transporter<\/jats:italic>\n              (PbCRT N331I) as responsible for this phenotype. In the current study, we generated a marker-free PbMut to enable further genetic manipulation of the isolated mutants. Here, we screened again for PPQ-resistant mutants in marker-free PbMut and obtained a parasite population with reduced susceptibility to PPQ. Of five isolated clones, none had the mutation PbCRT N331I; rather, they possessed a nonsense mutation at amino acid 119 (PbCRT Y119*), which would truncate the protein before eight of its ten predicted transmembrane domains. The PbCRT orthologue in the human malaria parasite\n              <jats:italic>Plasmodium falciparum<\/jats:italic>\n              , PfCRT, is an essential membrane transporter. To address the essentiality of PbCRT, we successfully deleted the full\n              <jats:italic>PbCRT<\/jats:italic>\n              gene [\n              <jats:italic>PbCRT<\/jats:italic>\n              (\u2212)] from wild-type parasites.\n              <jats:italic>PbCRT<\/jats:italic>\n              (\u2212) parasites exhibited reduced susceptibility to PPQ, along with compromised fitness in mice and following transmission to mosquitoes. Taken together, our findings provide the first evidence that\n              <jats:italic>P. berghei<\/jats:italic>\n              can acquire reduced PPQ susceptibility through complete loss of PbCRT function.\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1128\/aac.01589-24","type":"journal-article","created":{"date-parts":[[2025,2,24]],"date-time":"2025-02-24T14:00:24Z","timestamp":1740405624000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Deletion of the\n            <i>chloroquine resistance transporter<\/i>\n            gene confers reduced piperaquine susceptibility to the rodent malaria parasite\n            <i>Plasmodium berghei<\/i>"],"prefix":"10.1128","volume":"69","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5001-9653","authenticated-orcid":true,"given":"Makoto","family":"Hirai","sequence":"first","affiliation":[{"name":"Department of Tropical Medicine and Parasitology, Faculty of Medicine, Juntendo University","place":["Bunkyo-ku Hongo, Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meiji","family":"Arai","sequence":"additional","affiliation":[{"name":"Department of International Medical Zoology, School of Medicine, Kagawa University","place":["Kida, Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Soki","family":"Hayamichi","sequence":"additional","affiliation":[{"name":"Department of Tropical Medicine and Parasitology, Faculty of Medicine, Juntendo University","place":["Bunkyo-ku Hongo, Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ayako","family":"Uchida","sequence":"additional","affiliation":[{"name":"Department of Tropical Medicine and Parasitology, Faculty of Medicine, Juntendo University","place":["Bunkyo-ku Hongo, Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Megumi","family":"Sudo","sequence":"additional","affiliation":[{"name":"Department of Tropical Medicine and Parasitology, Faculty of Medicine, Juntendo University","place":["Bunkyo-ku Hongo, Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rie","family":"Kubota","sequence":"additional","affiliation":[{"name":"Department of Parasitology and Tropical Medicine, Graduate School of Medical and Dental Sciences, Institute of Science Tokyo","place":["Bunkyo-ku Yushima, Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Naoaki","family":"Shinzawa","sequence":"additional","affiliation":[{"name":"Department of Parasitology and Tropical Medicine, Graduate School of Medical and Dental Sciences, Institute of Science Tokyo","place":["Bunkyo-ku Yushima, Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toshihiro","family":"Mita","sequence":"additional","affiliation":[{"name":"Department of Tropical Medicine and Parasitology, Faculty of Medicine, Juntendo University","place":["Bunkyo-ku Hongo, Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_5_2_2","unstructured":"World Health Organization Geneva. 2023. 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