{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T05:53:57Z","timestamp":1777614837286,"version":"3.51.4"},"reference-count":11,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2024,11,5]],"date-time":"2024-11-05T00:00:00Z","timestamp":1730764800000},"content-version":"vor","delay-in-days":4,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001843","name":"Science and Engineering Research Board","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001843","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Reprod Domestic Animals"],"published-print":{"date-parts":[[2024,11]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>In mammals, sex determination is governed by the SRY gene on the Y chromosome, redirecting gonadal development from forming ovaries to testes. Mutations or alterations in the SRY gene can significantly affect phenotypic changes and lineage\u2010specific markers. This study aims to elucidate the role of the SRY gene in buffalo embryos using CRISPR\u2010Cas9 technology. We designed a crRNA targeting the HMG domain of the SRY gene using the CRISPOR algorithm. Nucleofection of sgRNA\u2010Cas9 RNPs into buffalo fibroblasts confirmed efficient cleavage at the targeted site. Using this validated guide, we investigated the role of the SRY gene in sexual determination by electroporating CRISPR\u2010Cas9\u2010RNPs into single\u2010stage zygotes of buffalo. Genetic changes in the SRY gene were confirmed through sequencing, revealing mosaic blastocysts with multiple alleles and non\u2010mosaic mutants. Mutations in SRY gene increased the expression of female lineage\u2010specific gene Wnt4 whereas decreased the expression of male specific gene SOX9 in blastocysts, suggesting reprogramming towards female sex determination pathways. Our findings provide insights into buffalo sex differentiation mechanisms and potential applications in reproductive strategies for breeding programmes.<\/jats:p>","DOI":"10.1111\/rda.14739","type":"journal-article","created":{"date-parts":[[2024,11,5]],"date-time":"2024-11-05T12:45:14Z","timestamp":1730810714000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["CRISPR\u2010Mediated SRY Gene Mutation Increases the Expression of Female Lineage\u2010Specific Gene in Pre\u2010Implantation Buffalo Embryo"],"prefix":"10.1111","volume":"59","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2171-8356","authenticated-orcid":false,"given":"Meeti","family":"Punetha","sequence":"first","affiliation":[{"name":"Animal Physiology and Reproduction Division ICAR\u2010Central Institute for Research on Buffaloes  Hisar India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sheetal","family":"Saini","sequence":"additional","affiliation":[{"name":"Animal Physiology and Reproduction Division ICAR\u2010Central Institute for Research on Buffaloes  Hisar India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Surabhi","family":"Sharma","sequence":"additional","affiliation":[{"name":"Animal Physiology and Reproduction Division ICAR\u2010Central Institute for Research on Buffaloes  Hisar India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Swati","family":"Thakur","sequence":"additional","affiliation":[{"name":"Animal Physiology and Reproduction Division ICAR\u2010Central Institute for Research on Buffaloes  Hisar India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Priya","family":"Dahiya","sequence":"additional","affiliation":[{"name":"Animal Physiology and Reproduction Division ICAR\u2010Central Institute for Research on Buffaloes  Hisar India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manu","family":"Mangal","sequence":"additional","affiliation":[{"name":"Animal Physiology and Reproduction Division ICAR\u2010Central Institute for Research on Buffaloes  Hisar India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rajesh","family":"Kumar","sequence":"additional","affiliation":[{"name":"Animal Physiology and Reproduction Division ICAR\u2010Central Institute for Research on Buffaloes  Hisar India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0521-8960","authenticated-orcid":false,"given":"Dharmendra","family":"Kumar","sequence":"additional","affiliation":[{"name":"Animal Physiology and Reproduction Division ICAR\u2010Central Institute for Research on Buffaloes  Hisar India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"P.\u00a0S.","family":"Yadav","sequence":"additional","affiliation":[{"name":"Animal Physiology and Reproduction Division ICAR\u2010Central Institute for Research on Buffaloes  Hisar India"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,11,5]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1210\/er.2002\u20100025"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-4658.2011.08029.x"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep03136"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/jez.1089"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2008743118"},{"issue":"8","key":"e_1_2_7_7_1","first-page":"863","article-title":"Conservation of SRY Function in Developing Testes of Mammals","volume":"36","author":"Lambeth L. S.","year":"2014","journal-title":"BioEssays"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2014.06.055"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.3390\/ani14010134"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tig.2008.10.008"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.18632\/oncotarget.16979"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1262\/jrd.2019-088"}],"container-title":["Reproduction in Domestic Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/rda.14739","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,11,27]],"date-time":"2024-11-27T10:50:17Z","timestamp":1732704617000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/rda.14739"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,11]]},"references-count":11,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2024,11]]}},"alternative-id":["10.1111\/rda.14739"],"URL":"https:\/\/doi.org\/10.1111\/rda.14739","archive":["Portico"],"relation":{},"ISSN":["0936-6768","1439-0531"],"issn-type":[{"value":"0936-6768","type":"print"},{"value":"1439-0531","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,11]]},"assertion":[{"value":"2024-08-09","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-10-15","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-11-05","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e14739"}}