{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T05:00:59Z","timestamp":1772168459112,"version":"3.50.1"},"reference-count":56,"publisher":"The Company of Biologists","issue":"9","license":[{"start":{"date-parts":[[2024,9,3]],"date-time":"2024-09-03T00:00:00Z","timestamp":1725321600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"NIH","award":["R01 GM134069"],"award-info":[{"award-number":["R01 GM134069"]}]},{"DOI":"10.13039\/501100004543","name":"China Scholarship Council","doi-asserted-by":"crossref","award":["201808210354"],"award-info":[{"award-number":["201808210354"]}],"id":[{"id":"10.13039\/501100004543","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Harvard Medical School; University of Utah"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,9,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Here, we introduce \u2018TICIT\u2019, targeted integration by CRISPR-Cas9 and integrase technologies, which utilizes the site-specific DNA recombinase \u2013 phiC31 integrase \u2013 to insert large DNA fragments into CRISPR-Cas9 target loci. This technique, which relies on first knocking in a 39-basepair phiC31 landing site via CRISPR-Cas9, enables researchers to repeatedly perform site-specific transgenesis at the exact genomic location with high precision and efficiency. We applied this approach to devise a method for the instantaneous determination of a zebrafish's genotype simply by examining its color. When a zebrafish mutant line must be propagated as heterozygotes due to homozygous lethality, employing this method allows facile identification of a population of homozygous mutant embryos even before the mutant phenotypes manifest. Thus, it should facilitate various downstream applications, such as large-scale chemical screens. We demonstrated that TICIT could also create reporter fish driven by an endogenous promoter. Further, we identified a landing site in the tyrosinase gene that could support transgene expression in a broad spectrum of tissue and cell types. In sum, TICIT enables site-specific DNA integration without requiring complex donor DNA construction. It can yield consistent transgene expression, facilitate diverse applications in zebrafish, and may be applicable to cells in culture and other model organisms.<\/jats:p>","DOI":"10.1242\/bio.061666","type":"journal-article","created":{"date-parts":[[2024,9,3]],"date-time":"2024-09-03T04:24:18Z","timestamp":1725337458000},"source":"Crossref","is-referenced-by-count":2,"title":["Instantaneous visual genotyping and facile site-specific transgenesis via CRISPR-Cas9 and phiC31 integrase"],"prefix":"10.1242","volume":"13","author":[{"given":"Junyan","family":"Ma","sequence":"first","affiliation":[{"name":"1 Department of Basic Medical Science, Quanzhou Medical College, Quanzhou, Fujian 362011, China"},{"name":"2 Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, MA 02129, USA and Department of Medicine, Harvard Medical School, Boston, MA 02115, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Weiting","family":"Zhang","sequence":"additional","affiliation":[{"name":"2 Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, MA 02129, USA and Department of Medicine, Harvard Medical School, Boston, MA 02115, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Simin","family":"Rahimialiabadi","sequence":"additional","affiliation":[{"name":"3 Department of Pharmacology and Toxicology, University of Utah, Salt Lake City, UT 84112, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nikkitha Umesh","family":"Ganesh","sequence":"additional","affiliation":[{"name":"2 Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, MA 02129, USA and Department of Medicine, Harvard Medical School, Boston, MA 02115, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhengwang","family":"Sun","sequence":"additional","affiliation":[{"name":"4 Center for Immunology and Inflammatory Disease, Massachusetts General Hospital, Charlestown, MA 02129, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Saba","family":"Parvez","sequence":"additional","affiliation":[{"name":"3 Department of Pharmacology and Toxicology, University of Utah, Salt Lake City, UT 84112, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Randall T.","family":"Peterson","sequence":"additional","affiliation":[{"name":"3 Department of Pharmacology and Toxicology, University of Utah, Salt Lake City, UT 84112, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7768-4901","authenticated-orcid":false,"given":"Jing-Ruey Joanna","family":"Yeh","sequence":"additional","affiliation":[{"name":"2 Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, MA 02129, USA and Department of Medicine, Harvard Medical School, Boston, MA 02115, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"237","published-online":{"date-parts":[[2024,9,3]]},"reference":[{"key":"2026022504473111600_BIO061666C1","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1016\/B978-0-12-374814-0.00002-1","article-title":"Tol2-mediated transgenesis, gene trapping, enhancer trapping, and the Gal4-UAS system","volume":"104","author":"Abe","year":"2011","journal-title":"Methods Cell Biol."},{"key":"2026022504473111600_BIO061666C2","doi-asserted-by":"publisher","first-page":"824","DOI":"10.1038\/s41587-020-0561-9","article-title":"Genome editing with CRISPR-Cas nucleases, base editors, transposases and prime editors","volume":"38","author":"Anzalone","year":"2020","journal-title":"Nat. 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