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Fluorescent protein tagging of endogenous protein in brain neurons using CRISPR/Cas9-mediated knock-in and in utero electroporation techniques
http://hdl.handle.net/10091/00021011
http://hdl.handle.net/10091/000210116a053a02-cefd-4969-84df-e68afdb8e0ac
| 名前 / ファイル | ライセンス | アクション |
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srep35861.pdf (2.4 MB)
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| Item type | 学術雑誌論文 / Journal Article(1) | |||||||||||||||||||||||||||||
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| 公開日 | 2018-10-31 | |||||||||||||||||||||||||||||
| タイトル | ||||||||||||||||||||||||||||||
| タイトル | Fluorescent protein tagging of endogenous protein in brain neurons using CRISPR/Cas9-mediated knock-in and in utero electroporation techniques | |||||||||||||||||||||||||||||
| 言語 | ||||||||||||||||||||||||||||||
| 言語 | eng | |||||||||||||||||||||||||||||
| DOI | ||||||||||||||||||||||||||||||
| 関連識別子 | https://doi.org/10.1038/srep35861 | |||||||||||||||||||||||||||||
| 関連名称 | 10.1038/srep35861 | |||||||||||||||||||||||||||||
| 資源タイプ | ||||||||||||||||||||||||||||||
| 資源 | http://purl.org/coar/resource_type/c_6501 | |||||||||||||||||||||||||||||
| タイプ | journal article | |||||||||||||||||||||||||||||
| 著者 |
Uemura, Takeshi
× Uemura, Takeshi
× Mori, Takuma
× Kurihara, Taiga
× Kawase, Shiori
× Koike, Rie
× Satoga, Michiru
× Cao, Xueshan
× Li, Xue
× Yanagawa, Toru
× Sakurai, Takayuki
× Shindo, Takayuki
× Tabuchi, Katsuhiko
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| 信州大学研究者総覧へのリンク | ||||||||||||||||||||||||||||||
| 氏名 | Uemura, Takeshi | |||||||||||||||||||||||||||||
| URL | http://soar-rd.shinshu-u.ac.jp/profile/ja.HmANWayV.html | |||||||||||||||||||||||||||||
| 出版者 | ||||||||||||||||||||||||||||||
| 出版者 | NATURE PUBLISHING GROUP | |||||||||||||||||||||||||||||
| 引用 | ||||||||||||||||||||||||||||||
| 内容記述 | SCIENTIFIC REPORTS.6:35861(2016) | |||||||||||||||||||||||||||||
| 書誌情報 |
SCIENTIFIC REPORTS 巻 6, p. 35861, 発行日 2016-10-26 |
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| 抄録 | ||||||||||||||||||||||||||||||
| 内容記述 | Genome editing is a powerful technique for studying gene functions. CRISPR/Cas9-mediated gene knock-in has recently been applied to various cells and organisms. Here, we successfully knocked in an EGFP coding sequence at the site immediately after the first ATG codon of the β-actin gene in neurons in the brain by the combined use of the CRISPR/Cas9 system and in utero electroporation technique, resulting in the expression of the EGFP-tagged β-actin protein in cortical layer 2/3 pyramidal neurons. We detected EGFP fluorescence signals in the soma and neurites of EGFP knock-in neurons. These signals were particularly abundant in the head of dendritic spines, corresponding to the localization of the endogenous β-actin protein. EGFP knock-in neurons showed no detectable changes in spine density and basic electrophysiological properties. In contrast, exogenously overexpressed EGFP-β-actin showed increased spine density and EPSC frequency, and changed resting membrane potential. Thus, our technique provides a potential tool to elucidate the localization of various endogenous proteins in neurons by epitope tagging without altering neuronal and synaptic functions. This technique can be also useful for introducing a specific mutation into genes to study the function of proteins and genomic elements in brain neurons. | |||||||||||||||||||||||||||||
| 資源タイプ（コンテンツの種類） | ||||||||||||||||||||||||||||||
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| 収録物識別子タイプ | EISSN | |||||||||||||||||||||||||||||
| 収録物識別子 | 2045-2322 | |||||||||||||||||||||||||||||
| PubMed | ||||||||||||||||||||||||||||||
| 識別子タイプ | PMID | |||||||||||||||||||||||||||||
| 関連識別子 | https://www.ncbi.nlm.nih.gov/pubmed/27782168 | |||||||||||||||||||||||||||||
| 関連名称 | 27782168 | |||||||||||||||||||||||||||||
| 権利 | ||||||||||||||||||||||||||||||
| 権利情報 | © The Author(s) 2016. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ | |||||||||||||||||||||||||||||
| 出版タイプ | ||||||||||||||||||||||||||||||
| 出版タイプ | VoR | |||||||||||||||||||||||||||||
| 出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||||||||||||||||||||||||||
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| URL | http://gateway.isiknowledge.com/gateway/Gateway.cgi?&GWVersion=2&SrcAuth=ShinshuUniv&SrcApp=ShinshuUniv&DestLinkType=FullRecord&DestApp=WOS&KeyUT=000386110600001 | |||||||||||||||||||||||||||||
