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CRISPR/Cas9-mediated gene knockout in the mouse brain using in utero electroporation
https://doi.org/10.24517/00014341
https://doi.org/10.24517/000143418f3537b0-16d8-4439-b4da-643fd64d3dce
| 名前 / ファイル | ライセンス | アクション |
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ME-PR-KAWASAKI-H-20611.pdf (726.7 kB)
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| Item type | 学術雑誌論文 / Journal Article(1) | |||||||||||||
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| 公開日 | 2017-10-03 | |||||||||||||
| タイトル | ||||||||||||||
| タイトル | CRISPR/Cas9-mediated gene knockout in the mouse brain using in utero electroporation | |||||||||||||
| 言語 | ||||||||||||||
| 言語 | eng | |||||||||||||
| 資源タイプ | ||||||||||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||||||||||
| 資源タイプ | journal article | |||||||||||||
| ID登録 | ||||||||||||||
| ID登録 | 10.24517/00014341 | |||||||||||||
| ID登録タイプ | JaLC | |||||||||||||
| 著者 |
Shinmyo, Yohei
× Shinmyo, Yohei× Tanaka, Satoshi× Tsunoda, Shinichi× Hosomichi, Kazuyoshi× Tajima, Atsushi× Kawasaki, Hiroshi |
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| 著者別表示 |
新明, 洋平
× 新明, 洋平
× 細道, 一善
× 田嶋, 敦
× 河崎, 洋志
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| 書誌情報 |
Scientific Reports 巻 6, p. 20611, 発行日 2016-02-09 |
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| ISSN | ||||||||||||||
| 収録物識別子タイプ | ISSN | |||||||||||||
| 収録物識別子 | 2045-2322 | |||||||||||||
| DOI | ||||||||||||||
| 関連タイプ | isIdenticalTo | |||||||||||||
| 識別子タイプ | DOI | |||||||||||||
| 関連識別子 | 10.1038/srep20611 | |||||||||||||
| 出版者 | ||||||||||||||
| 出版者 | Nature Publishing Group | |||||||||||||
| 抄録 | ||||||||||||||
| 内容記述タイプ | Abstract | |||||||||||||
| 内容記述 | 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. | |||||||||||||
| 著者版フラグ | ||||||||||||||
| 出版タイプ | VoR | |||||||||||||
| 出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||||||||||
