{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,30]],"date-time":"2026-06-30T01:24:49Z","timestamp":1782782689788,"version":"3.54.5"},"reference-count":50,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2022,2,15]],"date-time":"2022-02-15T00:00:00Z","timestamp":1644883200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Proyectos de Desarrollo Tecnol\u00f3gico AES 2019","award":["DTS19\/00061"],"award-info":[{"award-number":["DTS19\/00061"]}]},{"name":"MINECO-FEDER","award":["AGL 2015-66341-R"],"award-info":[{"award-number":["AGL 2015-66341-R"]}]},{"DOI":"10.13039\/100007801","name":"Fundaci\u00f3n Seneca","doi-asserted-by":"publisher","award":["20040\/GERM\/16"],"award-info":[{"award-number":["20040\/GERM\/16"]}],"id":[{"id":"10.13039\/100007801","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007801","name":"Fundaci\u00f3n Seneca","doi-asserted-by":"publisher","award":["21105\/PDC\/19"],"award-info":[{"award-number":["21105\/PDC\/19"]}],"id":[{"id":"10.13039\/100007801","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007801","name":"Fundaci\u00f3n Seneca","doi-asserted-by":"publisher","award":["21666\/PDC\/21"],"award-info":[{"award-number":["21666\/PDC\/21"]}],"id":[{"id":"10.13039\/100007801","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Spanish Ministry of Education, Culture and Sport","award":["FPU16\/04480"],"award-info":[{"award-number":["FPU16\/04480"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Mosaicism is the most important limitation for one-step gene editing in embryos by CRISPR\/Cas9 because cuts and repairs sometimes take place after the first DNA replication of the zygote. To try to minimize the risk of mosaicism, in this study a reversible DNA replication inhibitor was used after the release of CRISPR\/Cas9 in the cell. There is no previous information on the use of aphidicolin in porcine embryos, so the reversible inhibition of DNA replication and the effect on embryo development of different concentrations of this drug was first evaluated. The effect of incubation with aphidicolin was tested with CRISPR\/Cas9 at different concentrations and different delivery methodologies. As a result, the reversible inhibition of DNA replication was observed, and it was concentration dependent. An optimal concentration of 0.5 \u03bcM was established and used for subsequent experiments. Following the use of this drug with CRISPR\/Cas9, a halving of mosaicism was observed together with a detrimental effect on embryo development. In conclusion, the use of reversible inhibition of DNA replication offers a way to reduce mosaicism. Nevertheless, due to the reduction in embryo development, it would be necessary to reach a balance for its use to be feasible.<\/jats:p>","DOI":"10.3390\/ijms23042135","type":"journal-article","created":{"date-parts":[[2022,2,15]],"date-time":"2022-02-15T22:44:47Z","timestamp":1644965087000},"page":"2135","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Effect of Aphidicolin, a Reversible Inhibitor of Eukaryotic Nuclear DNA Replication, on the Production of Genetically Modified Porcine Embryos by CRISPR\/Cas9"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5210-1415","authenticated-orcid":false,"given":"Sergio","family":"Navarro-Serna","sequence":"first","affiliation":[{"name":"Department of Physiology, International Excellence Campus for Higher Education and Research \u201cCampus Mare Nostrum\u201d, University of Murcia, 30100 Murcia, Spain"},{"name":"Institute for Biomedical Research of Murcia (IMIB-Arrixaca), 30120 Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6139-2704","authenticated-orcid":false,"given":"Celia","family":"Pi\u00f1eiro-Silva","sequence":"additional","affiliation":[{"name":"Department of Physiology, International Excellence Campus for Higher Education and Research \u201cCampus Mare Nostrum\u201d, University of Murcia, 30100 Murcia, Spain"},{"name":"Institute for Biomedical Research of Murcia (IMIB-Arrixaca), 30120 Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chiara","family":"Luongo","sequence":"additional","affiliation":[{"name":"Department of Physiology, International Excellence Campus for Higher Education and Research \u201cCampus Mare Nostrum\u201d, University of Murcia, 30100 Murcia, Spain"},{"name":"Institute for Biomedical Research of Murcia (IMIB-Arrixaca), 30120 Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Parrington","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Oxford, Oxford OX1 3QT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0492-289X","authenticated-orcid":false,"given":"Raquel","family":"Romar","sequence":"additional","affiliation":[{"name":"Department of Physiology, International Excellence Campus for Higher Education and Research \u201cCampus Mare Nostrum\u201d, University of Murcia, 30100 Murcia, Spain"},{"name":"Institute for Biomedical Research of Murcia (IMIB-Arrixaca), 30120 Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3474-7626","authenticated-orcid":false,"given":"Joaqu\u00edn","family":"Gadea","sequence":"additional","affiliation":[{"name":"Department of Physiology, International Excellence Campus for Higher Education and Research \u201cCampus Mare Nostrum\u201d, University of Murcia, 30100 Murcia, Spain"},{"name":"Institute for Biomedical Research of Murcia (IMIB-Arrixaca), 30120 Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"7380","DOI":"10.1073\/pnas.77.12.7380","article-title":"Genetic transformation of mouse embryos by microinjection of purified DNA","volume":"77","author":"Gordon","year":"1980","journal-title":"Proc. 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