{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T10:13:34Z","timestamp":1789380814651,"version":"build-2803163510"},"reference-count":26,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2014,5,22]],"date-time":"2014-05-22T00:00:00Z","timestamp":1400716800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes to Cells"],"published-print":{"date-parts":[[2014,7]]},"abstract":"<jats:p>The type II clustered regularly interspaced short palindromic repeats (CRISPR)\/CRISPR\u2010associated (Cas) system, which is an adaptive immune system of bacteria, has become a powerful tool for genome editing in various model organisms. Here, we demonstrate multiple genome modifications mediated by CRISPR\/Cas9 in zebrafish (<jats:italic>Danio rerio<\/jats:italic>). Multiple genes including <jats:italic>golden\/gol<\/jats:italic> and <jats:italic>tyrosinase\/tyr<\/jats:italic>, which are involved in pigment formation, and <jats:italic>s1pr2<\/jats:italic> and <jats:italic>spns2<\/jats:italic>, which are involved in cardiac development, were disrupted with insertion and\/or deletion (indel) mutations introduced by the co\u2010injection of multiple guide RNAs (g<jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content>s) and the nuclease Cas9 m<jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content>. We simultaneously observed two distinct phenotypes, such as, the two hearts phenotype and the hypopigmentation of skin melanophores and the retinal pigment epithelium, in the injected F0 embryos. Additionally, we detected the targeted deletion and inversion genes as a 7.1\u2010kb fragment between the two distinct <jats:italic>spns2<\/jats:italic> targeted sites together with indel mutations. Conversely, chromosomal translocations among five target loci were not detected. Therefore, we confirmed that the CRISPR\/Cas9\u2010induced indel mutations and a locus\u2010specific deletion were heritable in F1 embryos. To screen founders, we improved heteroduplex mobility assay (HMA) for simultaneously detecting indel mutations in different target loci. The results suggest that the multi\u2010locus HMA is a powerful tool for identification of multiple genome modifications mediated by the CRISPR\/Cas9 system.<\/jats:p>","DOI":"10.1111\/gtc.12154","type":"journal-article","created":{"date-parts":[[2014,5,22]],"date-time":"2014-05-22T09:39:06Z","timestamp":1400751546000},"page":"555-564","source":"Crossref","is-referenced-by-count":110,"title":["Multiple genome modifications by the <scp>CRISPR<\/scp>\/Cas9 system in zebrafish"],"prefix":"10.1111","volume":"19","author":[{"given":"Satoshi","family":"Ota","sequence":"first","affiliation":[{"name":"Laboratory for Cardiovascular Molecular Dynamics RIKEN Quantitative Biology Center (QBiC) Furuedai 6\u20102\u20103 Suita Osaka 565\u20100874 Japan"},{"name":"Laboratory for Developmental Biology Center for Medical Education and Sciences Graduate School of Medical Science University of Yamanashi Shimogatou 1110 Chuo Yamanashi 409\u20103898 Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Hisano","sequence":"additional","affiliation":[{"name":"Laboratory for Cardiovascular Molecular Dynamics RIKEN Quantitative Biology Center (QBiC) Furuedai 6\u20102\u20103 Suita Osaka 565\u20100874 Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshiya","family":"Ikawa","sequence":"additional","affiliation":[{"name":"Department of Chemistry Graduate School of Science and Engineering University of Toyama Gofuku 3190 Toyama 930\u20108555 Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Atsuo","family":"Kawahara","sequence":"additional","affiliation":[{"name":"Laboratory for Cardiovascular Molecular Dynamics RIKEN Quantitative Biology Center (QBiC) Furuedai 6\u20102\u20103 Suita Osaka 565\u20100874 Japan"},{"name":"Laboratory for Developmental Biology Center for Medical Education and Sciences Graduate School of Medical Science University of Yamanashi Shimogatou 1110 Chuo Yamanashi 409\u20103898 Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2014,5,22]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btu048"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature11537"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1204094"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/cr.2013.45"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1101\/gr.162339.113"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1231143"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.1002861"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/dgd.12094"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.099085"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2501"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1308335110"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1225829"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1167449"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/35018092"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1116238"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2661"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2652"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1232033"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1319"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1111\/gtc.12050"},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/nn1344"},{"key":"e_1_2_6_23_1","doi-asserted-by":"publisher","DOI":"10.1101\/gr.163394.113"},{"key":"e_1_2_6_24_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrg2842"},{"key":"e_1_2_6_25_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature10886"},{"key":"e_1_2_6_26_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkt464"},{"key":"e_1_2_6_27_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2374"}],"container-title":["Genes 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Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fgtc.12154","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/gtc.12154","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,3]],"date-time":"2023-10-03T00:37:00Z","timestamp":1696293420000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/gtc.12154"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,5,22]]},"references-count":26,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2014,7]]}},"alternative-id":["10.1111\/gtc.12154"],"URL":"https:\/\/doi.org\/10.1111\/gtc.12154","archive":["Portico"],"relation":{},"ISSN":["1356-9597","1365-2443"],"issn-type":[{"value":"1356-9597","type":"print"},{"value":"1365-2443","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,5,22]]}}}