{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,5,29]],"date-time":"2025-05-29T04:12:19Z","timestamp":1748491939591,"version":"3.41.0"},"reference-count":33,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2025,5,28]],"date-time":"2025-05-28T00:00:00Z","timestamp":1748390400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Key Research and Development Program of China","award":["2023YFD1801000"],"award-info":[{"award-number":["2023YFD1801000"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["U21A20258"],"award-info":[{"award-number":["U21A20258"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100021171","name":"Basic and Applied Basic Research Foundation of Guangdong Province","doi-asserted-by":"publisher","award":["2023A1515012862"],"award-info":[{"award-number":["2023A1515012862"]}],"id":[{"id":"10.13039\/501100021171","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Double First-Class Discipline Promotion Project","award":["2023B10564003"],"award-info":[{"award-number":["2023B10564003"]}]},{"DOI":"10.13039\/501100013314","name":"111 Project","doi-asserted-by":"crossref","award":["D20008"],"award-info":[{"award-number":["D20008"]}],"id":[{"id":"10.13039\/501100013314","id-type":"DOI","asserted-by":"crossref"}]},{"name":"The Yunnan Key Laboratory of Veterinary Etiological Biology","award":["202449CE340019"],"award-info":[{"award-number":["202449CE340019"]}]},{"name":"Yunnan Agricultural Joint Special Project","award":["202301BD070001-183"],"award-info":[{"award-number":["202301BD070001-183"]}]},{"name":"Key Laboratory of Veterinary Etiological Biology, Ministry of Agriculture and Rural Affairs, China","award":["XKQ2024005"],"award-info":[{"award-number":["XKQ2024005"]}]}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"<jats:sec id=\"sec016\">\n<jats:title>Background<\/jats:title>\n<jats:p><jats:italic>Cryptosporidium parvum<\/jats:italic> is a protozoan pathogen that causes moderate to severe diarrhea in both humans and animals. Calcium-dependent protein kinases (CDPKs) are attractive drug targets against cryptosporidiosis given their critical role in the life cycle of <jats:italic>Cryptosporidium<\/jats:italic> spp. and their absence in human and animal hosts.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec017\">\n<jats:title>Methodology\/principal findings<\/jats:title>\n<jats:p>We used CRISPR-Cas9 technology to endogenously tag the <jats:italic>CpCDPK2A<\/jats:italic> gene in <jats:italic>C. parvum<\/jats:italic> IIdA20G1-HLJ strain with the hemagglutinin (HA) epitope and to delete the <jats:italic>CpCDPK2A<\/jats:italic> gene. An immunofluorescence assay was performed to localize the CpCDPK2A expression in the tagged strain and a luciferase assay was performed to compare growth rates of the tagged and deletion strains <jats:italic>in vitro<\/jats:italic>. Oocyst shedding, parasite load, villus length\/crypt height ratio and survival of infected mice were used to evaluate the function of CpCDPK2A <jats:italic>in vivo<\/jats:italic>. The results revealed that CpCDPK2A was expressed in all the intracellular developmental stages, especially in the motile stages of sporozoites and merozoites. While CpCDPK2A is dispensable, deletion of the gene significantly reduced the growth of late asexual and sexual stages <jats:italic>in vitro<\/jats:italic>. In an interferon-\u03b3 knockout mouse model, gene deletion of <jats:italic>CpCDPK2A<\/jats:italic> reduced oocyst shedding by 25-fold and increased survival of infected mice.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec018\">\n<jats:title>Conclusions\/significance<\/jats:title>\n<jats:p>These observations suggest that CpCDPK2A may contribute to both asexual and sexual replication of <jats:italic>C. parvum<\/jats:italic> and may be a potential target to block the transmission of this important zoonotic pathogen.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pntd.0013107","type":"journal-article","created":{"date-parts":[[2025,5,28]],"date-time":"2025-05-28T21:18:28Z","timestamp":1748467108000},"page":"e0013107","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Calcium-dependent protein kinases 2A involved in the growth of both asexual and sexual stages of Cryptosporidium parvum"],"prefix":"10.1371","volume":"19","author":[{"given":"Fanfan","family":"Shu","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yujin","family":"Huang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fuxian","family":"Yang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yaqiong","family":"Guo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rui","family":"Xu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lihua","family":"Xiao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yaoyu","family":"Feng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0114-4877","authenticated-orcid":true,"given":"Na","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2025,5,28]]},"reference":[{"issue":"1","key":"pntd.0013107.ref001","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/S1473-3099(14)70772-8","article-title":"A 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