{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,8,2]],"date-time":"2025-08-02T19:22:26Z","timestamp":1754162546650,"version":"3.41.2"},"reference-count":30,"publisher":"PeerJ","license":[{"start":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T00:00:00Z","timestamp":1753833600000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key R&D Program of China","doi-asserted-by":"crossref","award":["2022YFC2702800"],"award-info":[{"award-number":["2022YFC2702800"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:sec>\n               <jats:title>Background<\/jats:title>\n               <jats:p>Ring finger motifs are found in a variety of proteins with diverse functions, often involved in protein-DNA or protein\u2013protein interactions. The <jats:italic>Rnf32<\/jats:italic>-encoded protein contains two such motifs and is predominantly expressed in the testes and ovaries, suggesting that its expression may be regulated by elements within the <jats:italic>Rnf32<\/jats:italic> promoter region. <jats:italic>Rnf32<\/jats:italic> is active during spermatogenesis, mainly in spermatocytes and spermatids, indicating a potential role in sperm development.<\/jats:p>\n            <\/jats:sec>\n            <jats:sec>\n               <jats:title>Methods<\/jats:title>\n               <jats:p>We established an <jats:italic>Rnf32<\/jats:italic> knockout (<jats:italic>Rnf32<\/jats:italic><jats:sup>\u2212\/\u2212<\/jats:sup>) mouse model using CRISPR\/Cas9 technology. Gene expression was analyzed <jats:italic>via<\/jats:italic> reverse transcription quantitative polymerase chain reaction (RT-qPCR). Testicular and epididymal phenotypes were assessed through histological and immunofluorescence staining, and fertility and sperm motility were evaluated.<\/jats:p>\n            <\/jats:sec>\n            <jats:sec>\n               <jats:title>Results<\/jats:title>\n               <jats:p>Here, we successfully established an <jats:italic>Rnf32<\/jats:italic> knockout mouse model using CRISPR\/Cas9 technology. Surprisingly, male <jats:italic>Rnf32<\/jats:italic><jats:sup>\u2212\/\u2212<\/jats:sup> mice exhibited normal fertility, with no significant differences in testicular and epididymal histology, spermatogenesis, sperm count, or motility compared to <jats:italic>Rnf32<\/jats:italic><jats:sup>+\/+<\/jats:sup> mice. These findings suggest that <jats:italic>Rnf32<\/jats:italic> may not be essential for male fertility in mice, and its potential functions warrant further investigation.<\/jats:p>\n            <\/jats:sec>","DOI":"10.7717\/peerj.19794","type":"journal-article","created":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T08:46:22Z","timestamp":1753865182000},"page":"e19794","source":"Crossref","is-referenced-by-count":0,"title":["<i>Rnf32<\/i> is not essential for spermatogenesis and male fertility in mice"],"prefix":"10.7717","volume":"13","author":[{"given":"Hao","family":"Kong","sequence":"first","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yufeng","family":"Yin","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ni","family":"Zeng","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yunfei","family":"Zhu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yiqiang","family":"Cui","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"4443","published-online":{"date-parts":[[2025,7,30]]},"reference":[{"issue":"3","key":"10.7717\/peerj.19794\/ref-1","first-page":"286","article-title":"The cycle of the seminiferous epithelium and stages in spermatogenesis in dd-mice","volume":"66","author":"Abe","year":"1991","journal-title":"[Hokkaido Igaku Zasshi] The Hokkaido Journal of Medical Science"},{"key":"10.7717\/peerj.19794\/ref-2","doi-asserted-by":"publisher","first-page":"2487","DOI":"10.7150\/ijbs.60384","article-title":"Towards post-meiotic sperm production: genetic insight into human infertility from mouse models","volume":"17","author":"Azhar","year":"2021","journal-title":"International Journal of Biological Sciences"},{"key":"10.7717\/peerj.19794\/ref-3","doi-asserted-by":"publisher","first-page":"660","DOI":"10.1093\/humupd\/dmx021","article-title":"The diagnosis of male infertility: an analysis of the evidence to support the development of global WHO guidance-challenges and future research opportunities","volume":"23","author":"Barratt","year":"2017","journal-title":"Human Reproduction Update"},{"key":"10.7717\/peerj.19794\/ref-4","doi-asserted-by":"publisher","first-page":"3716","DOI":"10.2174\/138161209789271825","article-title":"RING finger E(3) ubiquitin ligases: structure and drug discovery","volume":"15","author":"Chasapis","year":"2009","journal-title":"Current Pharmaceutical Design"},{"key":"10.7717\/peerj.19794\/ref-5","doi-asserted-by":"publisher","first-page":"395","DOI":"10.1186\/s12864-024-10298-y","article-title":"Multiple transcriptome analyses reveal mouse testis developmental dynamics","volume":"25","author":"Chen","year":"2024","journal-title":"BMC Genomics"},{"key":"10.7717\/peerj.19794\/ref-6","doi-asserted-by":"publisher","first-page":"7287","DOI":"10.1007\/s11033-022-07512-z","article-title":"Fbxw17 is dispensable for viability and fertility in mice","volume":"49","author":"Chen","year":"2022","journal-title":"Molecular Biology Reports"},{"key":"10.7717\/peerj.19794\/ref-7","doi-asserted-by":"publisher","first-page":"879","DOI":"10.1038\/s41422-018-0074-y","article-title":"Single-cell RNA-seq uncovers dynamic processes and critical regulators in mouse spermatogenesis","volume":"28","author":"Chen","year":"2018","journal-title":"Cell Research"},{"key":"10.7717\/peerj.19794\/ref-8","doi-asserted-by":"publisher","first-page":"790","DOI":"10.1038\/nrg911","article-title":"Mouse models of male infertility","volume":"3","author":"Cooke","year":"2002","journal-title":"Nature Reviews. 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