{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:01:45Z","timestamp":1760144505775,"version":"build-2065373602"},"reference-count":27,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2024,4,23]],"date-time":"2024-04-23T00:00:00Z","timestamp":1713830400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["2021R1C1C1011346","2022R1A2C1004096","21C0726L1","HI22C1988"],"award-info":[{"award-number":["2021R1C1C1011346","2022R1A2C1004096","21C0726L1","HI22C1988"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Korean Fund for Regenerative Medicine","award":["2021R1C1C1011346","2022R1A2C1004096","21C0726L1","HI22C1988"],"award-info":[{"award-number":["2021R1C1C1011346","2022R1A2C1004096","21C0726L1","HI22C1988"]}]},{"name":"Korea Health Technology R&amp;D Project","award":["2021R1C1C1011346","2022R1A2C1004096","21C0726L1","HI22C1988"],"award-info":[{"award-number":["2021R1C1C1011346","2022R1A2C1004096","21C0726L1","HI22C1988"]}]},{"name":"Jeonbuk National University","award":["2021R1C1C1011346","2022R1A2C1004096","21C0726L1","HI22C1988"],"award-info":[{"award-number":["2021R1C1C1011346","2022R1A2C1004096","21C0726L1","HI22C1988"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["CIMB"],"abstract":"<jats:p>Rat animal models are widely used owing to their relatively superior cognitive abilities and higher similarity compared with mouse models to human physiological characteristics. However, their use is limited because of difficulties in establishing embryonic stem cells and performing genetic modifications, and insufficient embryological research. In this study, we established optimal superovulation and fertilized\u2013egg transfer conditions, including optimal hormone injection concentration (\u2265150 IU\/kg of PMSG and hCG) and culture medium (mR1ECM), to obtain high-quality zygotes and establish in vitro fertilization conditions for rats. Next, sgRNA with optimal targeting activity was selected by performing PCR analysis and the T7E1 assay, and the CRISPR\/Cas9 system was used to construct a rat model for muscular dystrophy by inducing a deficiency in the fukutin gene without any off-target effect detected. The production of fukutin knockout rats was phenotypically confirmed by observing a drop-in body weight to one-third of that of the control group. In summary, we succeeded in constructing the first muscular dystrophy disease rat model using the CRISPR\/CAS9 system for increasing future prospects of producing various animal disease models and encouraging disease research using rats.<\/jats:p>","DOI":"10.3390\/cimb46050234","type":"journal-article","created":{"date-parts":[[2024,4,23]],"date-time":"2024-04-23T05:28:06Z","timestamp":1713850086000},"page":"3752-3762","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Optimizing Embryo Collection for Application of CRISPR\/Cas9 System and Generation of Fukutin Knockout Rat Using This Method"],"prefix":"10.3390","volume":"46","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7400-9022","authenticated-orcid":false,"given":"Dong-Won","family":"Seol","sequence":"first","affiliation":[{"name":"Preclinical Research Center, Daegu-Gyeongbuk Medical Innovation Foundation (KMEDIHUB), Daegu 41061, Republic of Korea"},{"name":"Non-Clinical Evaluation Center, Osong Medical Innovation Foundation (KBIO Health), Cheongju 28160, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Byoung-Jin","family":"Park","sequence":"additional","affiliation":[{"name":"Primate Resources Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7825-9598","authenticated-orcid":false,"given":"Deog-Bon","family":"Koo","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Daegu University, Gyeongsan 38453, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ji-Su","family":"Kim","sequence":"additional","affiliation":[{"name":"Primate Resources Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yong-Hyun","family":"Jeon","sequence":"additional","affiliation":[{"name":"Preclinical Research Center, Daegu-Gyeongbuk Medical Innovation Foundation (KMEDIHUB), Daegu 41061, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jae-Eon","family":"Lee","sequence":"additional","affiliation":[{"name":"Preclinical Research Center, Daegu-Gyeongbuk Medical Innovation Foundation (KMEDIHUB), Daegu 41061, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joon-Suk","family":"Park","sequence":"additional","affiliation":[{"name":"Preclinical Research Center, Daegu-Gyeongbuk Medical Innovation Foundation (KMEDIHUB), Daegu 41061, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hoon","family":"Jang","sequence":"additional","affiliation":[{"name":"Department of Life Science, Jeonbuk National University, Jeonju 54896, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0201-0416","authenticated-orcid":false,"given":"Gabbine","family":"Wee","sequence":"additional","affiliation":[{"name":"Preclinical Research Center, Daegu-Gyeongbuk Medical Innovation Foundation (KMEDIHUB), Daegu 41061, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,4,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1177\/026119299802600108","article-title":"The Use of Transgenic Animals in the European Union: The Report and Recommendations of ECVAM Workshop 28","volume":"26","author":"Mepham","year":"1998","journal-title":"Altern. 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