| Item type |
学術雑誌論文 / Journal Article(1) |
| 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
Tanaka, Satoshi
Tsunoda, Shinichi
Hosomichi, Kazuyoshi
Tajima, Atsushi
Kawasaki, Hiroshi
|
| 著者別表示 |
新明, 洋平
細道, 一善
田嶋, 敦
河崎, 洋志
|
| 書誌情報 |
Scientific Reports
巻 6,
p. 20611,
発行日 2016-02-09
|
| 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 |
ME-PR-KAWASAKI-H-20611.pdf (726.7 kB)
.png)
