{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,17]],"date-time":"2026-03-17T21:46:39Z","timestamp":1773783999332,"version":"3.50.1"},"reference-count":23,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,10,9]],"date-time":"2021-10-09T00:00:00Z","timestamp":1633737600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,10,9]],"date-time":"2021-10-09T00:00:00Z","timestamp":1633737600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Res Notes"],"published-print":{"date-parts":[[2021,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec>\n                <jats:title>Objective<\/jats:title>\n                <jats:p>Lipofection-mediated introduction of the CRISPR\/Cas9 system in porcine zygotes provides a simple method for gene editing, without requiring micromanipulation. However, the gene editing efficiency is inadequate. The aim of this study was to improve the lipofection-mediated gene editing efficiency by optimizing the timing and duration of lipofection.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>Zona pellucida (ZP)-free zygotes collected at 5, 10, and 15\u00a0h from the start of in vitro fertilization (IVF) were incubated with lipofection reagent, guide RNA (gRNA) targeting <jats:italic>GGTA1<\/jats:italic>, and Cas9 for 5\u00a0h. Lipofection of zygotes collected at 10 and 15\u00a0h from the start of IVF yielded mutant blastocysts. Next, ZP-free zygotes collected at 10\u00a0h from the start of IVF were incubated with lipofection reagent, gRNA, and Cas9 for 2.5, 5, 10, or 20\u00a0h. The blastocyst formation rate of zygotes treated for 20\u00a0h was significantly lower (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.05) than those of the other groups, and no mutant blastocysts were obtained. Moreover, the mutation rates of the resulting blastocysts decreased as the incubation time increased. In conclusion, a lipofection-mediated gene editing system using the CRISPR\/Cas9 system in ZP-zygotes is feasible; however, further improvements in the gene editing efficiency are required.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1186\/s13104-021-05800-8","type":"journal-article","created":{"date-parts":[[2021,10,10]],"date-time":"2021-10-10T20:55:43Z","timestamp":1633899343000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Timing and duration of lipofection-mediated CRISPR\/Cas9 delivery into porcine zygotes affect gene-editing events"],"prefix":"10.1186","volume":"14","author":[{"given":"Qingyi","family":"Lin","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Quynh Anh","family":"Le","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Koki","family":"Takebayashi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chommanart","family":"Thongkittidilok","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manita","family":"Wittayarat","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maki","family":"Hirata","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2158-5808","authenticated-orcid":false,"given":"Fuminori","family":"Tanihara","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takeshige","family":"Otoi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2021,10,9]]},"reference":[{"key":"5800_CR1","doi-asserted-by":"publisher","first-page":"273","DOI":"10.1007\/s11248-016-9932-x","volume":"25","author":"W Tan","year":"2016","unstructured":"Tan W, Proudfoot C, Lillico SG, Whitelaw CB. 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