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Among the three <jats:italic>Cryptosporidium<\/jats:italic> species infecting the intestines of calves, <jats:italic>Cryptosporidium parvum<\/jats:italic> has a broad host range and causes severe diarrhea in calves, while <jats:italic>Cryptosporidium bovis<\/jats:italic> and <jats:italic>Cryptosporidium ryanae<\/jats:italic> mainly infect calves without obvious clinical symptoms. Comparative genomic analysis revealed differences in the copy number of genes encoding the nonfinancial disclosure quality (NFDQ) secretory protein family among the three species, suggesting that this protein family may be associated with the host range or pathogenicity of <jats:italic>Cryptosporidium<\/jats:italic> spp. To understand the function of cgd8_10 encoded NFDQ1, tagged and knockout strains were constructed and characterized in this study.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>To determine the localization of NFDQ1, we used clustered regularly interspaced short palindromic repeats (CRISPR)\/CRISPR-associated protein 9 (Cas9) technology to tag the C-terminus of NFDQ1 with three hemagglutinin epitopes (3\u2009\u00d7\u2009HA). The tagged strain was constructed, and the genomic insertion was confirmed by polymerase chain reaction (PCR). Immunofluorescence assays were performed to observe the localization of NFDQ1 both in extracellular sporozoites and at various intracellular developmental stages. Immunoelectron microscopy was used to study the ultrastructural localization of NFDQ1. Then, the <jats:italic>\u0394NFDQ1<\/jats:italic> strain was generated by CRISPR\/Cas9 and the in vitro growth assay on HCT-8 cells was used to analyze of phenotypic changes after knockout <jats:italic>NFDQ1<\/jats:italic> in parasites.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>The <jats:italic>NFDQ1<\/jats:italic> tagging and knockout stains were successfully constructed by CRISPR\/Cas9 technology and the insertions of transgenic strains were validated by PCR. The expression of NFDQ1 was validated in parasite by western blot. Immunofluorescence and immune-electron microscopy assay showed that NFDQ1 expressed in both asexual and sexual stages of <jats:italic>C<\/jats:italic>. <jats:italic>parvum<\/jats:italic>, where it was localized to the cytoplasm of the parasite. Upon ablation of <jats:italic>NFDQ1<\/jats:italic>, the <jats:italic>\u0394NFDQ1<\/jats:italic> strain showed an apparent growth retardation during sexual replication in vitro.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>NFDQ1 is a cytoplasmic protein without specific localization to secretory organelles, and it may participate in <jats:italic>C<\/jats:italic>. <jats:italic>parvum<\/jats:italic> growth during sexual reproduction. Future study should determine the role of NFDQ1 following <jats:italic>C<\/jats:italic>. <jats:italic>parvum<\/jats:italic> infection in vivo.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Graphical Abstract<\/jats:title>\n                \n              <\/jats:sec>","DOI":"10.1186\/s13071-024-06532-x","type":"journal-article","created":{"date-parts":[[2024,10,26]],"date-time":"2024-10-26T17:01:49Z","timestamp":1729962109000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Characterization of NFDQ1 in Cryptosporidium parvum"],"prefix":"10.1186","volume":"17","author":[{"given":"Yangsiqi","family":"Ao","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaoqing","family":"Gong","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jieping","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruiming","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shujiao","family":"Song","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yaqiong","family":"Guo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yaoyu","family":"Feng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lihua","family":"Xiao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rui","family":"Xu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Na","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,10,26]]},"reference":[{"key":"6532_CR1","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1038\/503189a","volume":"503","author":"B Striepen","year":"2013","unstructured":"Striepen B. 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