{"created":"2023-07-27T06:29:34.557084+00:00","id":14354,"links":{},"metadata":{"_buckets":{"deposit":"7793d757-8c65-46db-a741-9e99cff551fe"},"_deposit":{"created_by":3,"id":"14354","owners":[3],"pid":{"revision_id":0,"type":"depid","value":"14354"},"status":"published"},"_oai":{"id":"oai:kanazawa-u.repo.nii.ac.jp:00014354","sets":["4280:4292:4313"]},"author_link":["24834","190","24833","24832","69683","69684","95464","24011","24080"],"item_4_biblio_info_8":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2016-02-09","bibliographicIssueDateType":"Issued"},"bibliographicPageStart":"20611","bibliographicVolumeNumber":"6","bibliographic_titles":[{"bibliographic_title":"Scientific Reports"}]}]},"item_4_creator_33":{"attribute_name":"著者別表示","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"新明, 洋平"}],"nameIdentifiers":[{},{},{},{}]},{"creatorNames":[{"creatorName":"細道, 一善"}],"nameIdentifiers":[{},{}]},{"creatorNames":[{"creatorName":"田嶋, 敦"}],"nameIdentifiers":[{},{}]},{"creatorNames":[{"creatorName":"河崎, 洋志"}],"nameIdentifiers":[{},{}]}]},"item_4_description_21":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"The CRISPR/Cas9 system has recently been adapted for generating knockout mice to investigate physiological functions and pathological mechanisms. Here, we report a highly efficient procedure for brain-specific disruption of genes of interest in vivo. We constructed pX330 plasmids expressing humanized Cas9 and single-guide RNAs (sgRNAs) against the Satb2 gene, which encodes an AT-rich DNA-binding transcription factor and is responsible for callosal axon projections in the developing mouse brain. We first confirmed that these constructs efficiently induced double-strand breaks (DSBs) in target sites of exogenous plasmids both in vitro and in vivo. We then found that the introduction of pX330-Satb2 into the developing mouse brain using in utero electroporation led to a dramatic reduction of Satb2 expression in the transfected cerebral cortex, suggesting DSBs had occurred in the Satb2 gene with high efficiency. Furthermore, we found that Cas9-mediated targeting of the Satb2 gene induced abnormalities in axonal projection patterns, which is consistent with the phenotypes previously observed in Satb2 mutant mice. Introduction of pX330-NeuN using our procedure also resulted in the efficient disruption of the NeuN gene. Thus, our procedure combining the CRISPR/Cas9 system and in utero electroporation is an effective and rapid approach to achieve brain-specific gene knockout in vivo. © 2016, Nature Publishing Group. All rights reserved.","subitem_description_type":"Abstract"}]},"item_4_identifier_registration":{"attribute_name":"ID登録","attribute_value_mlt":[{"subitem_identifier_reg_text":"10.24517/00014341","subitem_identifier_reg_type":"JaLC"}]},"item_4_publisher_17":{"attribute_name":"出版者","attribute_value_mlt":[{"subitem_publisher":"Nature Publishing Group"}]},"item_4_relation_12":{"attribute_name":"DOI","attribute_value_mlt":[{"subitem_relation_type":"isIdenticalTo","subitem_relation_type_id":{"subitem_relation_type_id_text":"10.1038/srep20611","subitem_relation_type_select":"DOI"}}]},"item_4_source_id_9":{"attribute_name":"ISSN","attribute_value_mlt":[{"subitem_source_identifier":"2045-2322","subitem_source_identifier_type":"ISSN"}]},"item_4_version_type_25":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_970fb48d4fbd8a85","subitem_version_type":"VoR"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"Shinmyo, Yohei"}],"nameIdentifiers":[{},{},{},{}]},{"creatorNames":[{"creatorName":"Tanaka, Satoshi"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"Tsunoda, Shinichi"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"Hosomichi, Kazuyoshi"}],"nameIdentifiers":[{},{},{}]},{"creatorNames":[{"creatorName":"Tajima, Atsushi"}],"nameIdentifiers":[{},{},{},{}]},{"creatorNames":[{"creatorName":"Kawasaki, Hiroshi"}],"nameIdentifiers":[{},{},{},{}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2017-10-03"}],"displaytype":"detail","filename":"ME-PR-KAWASAKI-H-20611.pdf","filesize":[{"value":"726.7 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"ME-PR-KAWASAKI-H-20611.pdf","url":"https://kanazawa-u.repo.nii.ac.jp/record/14354/files/ME-PR-KAWASAKI-H-20611.pdf"},"version_id":"120835dd-0e76-4d6c-af97-54e135d1c533"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"eng"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"journal article","resourceuri":"http://purl.org/coar/resource_type/c_6501"}]},"item_title":"CRISPR/Cas9-mediated gene knockout in the mouse brain using in utero electroporation","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"CRISPR/Cas9-mediated gene knockout in the mouse brain using in utero electroporation"}]},"item_type_id":"4","owner":"3","path":["4313"],"pubdate":{"attribute_name":"公開日","attribute_value":"2017-10-03"},"publish_date":"2017-10-03","publish_status":"0","recid":"14354","relation_version_is_last":true,"title":["CRISPR/Cas9-mediated gene knockout in the mouse brain using in utero electroporation"],"weko_creator_id":"3","weko_shared_id":3},"updated":"2023-07-27T08:47:44.210873+00:00"}