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Here, we report a highly efficient procedure for brain-specific disruption of genes of interest\n                    <jats:italic>in vivo<\/jats:italic>\n                    . We constructed pX330 plasmids expressing humanized Cas9 and single-guide RNAs (sgRNAs) against the\n                    <jats:italic>Satb2<\/jats:italic>\n                    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\n                    <jats:italic>in vitro<\/jats:italic>\n                    and\n                    <jats:italic>in vivo<\/jats:italic>\n                    . We then found that the introduction of pX330-Satb2 into the developing mouse brain using\n                    <jats:italic>in utero<\/jats:italic>\n                    electroporation led to a dramatic reduction of Satb2 expression in the transfected cerebral cortex, suggesting DSBs had occurred in the\n                    <jats:italic>Satb2<\/jats:italic>\n                    gene with high efficiency. Furthermore, we found that Cas9-mediated targeting of the\n                    <jats:italic>Satb2<\/jats:italic>\n                    gene induced abnormalities in axonal projection patterns, which is consistent with the phenotypes previously observed in\n                    <jats:italic>Satb2<\/jats:italic>\n                    mutant mice. Introduction of pX330-NeuN using our procedure also resulted in the efficient disruption of the\n                    <jats:italic>NeuN<\/jats:italic>\n                    gene. Thus, our procedure combining the CRISPR\/Cas9 system and\n                    <jats:italic>in utero<\/jats:italic>\n                    electroporation is an effective and rapid approach to achieve brain-specific gene knockout\n                    <jats:italic>in vivo<\/jats:italic>\n                    .\n                  <\/jats:p>","DOI":"10.1038\/srep20611","type":"journal-article","created":{"date-parts":[[2016,2,9]],"date-time":"2016-02-09T05:39:52Z","timestamp":1454996392000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":80,"title":["CRISPR\/Cas9-mediated gene knockout in the mouse brain using in utero electroporation"],"prefix":"10.1038","volume":"6","author":[{"given":"Yohei","family":"Shinmyo","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Satoshi","family":"Tanaka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shinichi","family":"Tsunoda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazuyoshi","family":"Hosomichi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Atsushi","family":"Tajima","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroshi","family":"Kawasaki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2016,2,9]]},"reference":[{"key":"BFsrep20611_CR1","doi-asserted-by":"publisher","first-page":"507","DOI":"10.1038\/nrg1619","volume":"6","author":"MR Capecchi","year":"2005","unstructured":"Capecchi, M. 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