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Efficient Generation of Somatic Cell Nuclear Transfer-Competent Porcine Cells with Mutated Alleles at Multiple Target Loci by Using CRISPR/Cas9 Combined with Targeted Toxin-Based Selection System
http://hdl.handle.net/10091/00021603
http://hdl.handle.net/10091/000216035038cfa4-3c6f-4b85-8a7e-636eb806f017
名前 / ファイル | ライセンス | アクション |
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16K15233_04.pdf (25.5 MB)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2019-09-09 | |||||
タイトル | ||||||
タイトル | Efficient Generation of Somatic Cell Nuclear Transfer-Competent Porcine Cells with Mutated Alleles at Multiple Target Loci by Using CRISPR/Cas9 Combined with Targeted Toxin-Based Selection System | |||||
言語 | en | |||||
言語 | ||||||
言語 | eng | |||||
DOI | ||||||
識別子タイプ | DOI | |||||
関連識別子 | https://doi.org/10.3390/ijms18122610 | |||||
関連名称 | 10.3390/ijms18122610 | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | alpha-Gal epitope | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | alpha-1,3-galactosyltransferase | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | CRISPR/Cas9 | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | targeted toxin | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | genome editing | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | isolectin BS-I-B-4 | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | LDLR | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | PTEN | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | p53 | |||||
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 | |||||
著者 |
Sato, Masahiro
× Sato, Masahiro× Miyoshi, Kazuchika× Nakamura, Shingo× Ohtsuka, Masato× Sakurai, Takayuki× Watanabe, Satoshi× Kawaguchi, Hiroaki× Tanimoto, Akihide |
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信州大学研究者総覧へのリンク | ||||||
表示名 | Sakurai, Takayuki | |||||
URL | http://soar-rd.shinshu-u.ac.jp/profile/ja.jaLUuNkh.html | |||||
出版者 | ||||||
出版者 | MDPI | |||||
引用 | ||||||
内容記述タイプ | Other | |||||
内容記述 | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES.18(12):2610(2017) | |||||
書誌情報 |
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 巻 18, 号 12, p. 2610, 発行日 2017-12-04 |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | The recent advancement in genome editing such a CRISPR/Cas9 system has enabled isolation of cells with knocked multiple alleles through a one-step transfection. Somatic cell nuclear transfer (SCNT) has been frequently employed as one of the efficient tools for the production of genetically modified (GM) animals. To use GM cells as SCNT donor, efficient isolation of transfectants with mutations at multiple target loci is often required. The methods for the isolation of such GM cells largely rely on the use of drug selection-based approach using selectable genes; however, it is often difficult to isolate cells with mutations at multiple target loci. In this study, we used a novel approach for the efficient isolation of porcine cells with at least two target loci mutations by one-step introduction of CRISPR/Cas9-related components. A single guide (sg) RNA targeted to GGTA1 gene, involved in the synthesis of cell-surface alpha-Gal epitope (known as xenogenic antigen), is always a prerequisite. When the transfected cells were reacted with toxin-labeled BS-I-B-4 isolectin for 2 h at 37 degrees C to eliminate alpha-Gal epitope-expressing cells, the surviving clones lacked alpha-Gal epitope expression and were highly expected to exhibit induced mutations at another target loci. Analysis of these alpha-Gal epitope-negative surviving cells demonstrated a 100% occurrence of genome editing at target loci. SCNT using these cells as donors resulted in the production of cloned blastocysts with the genotype similar to that of the donor cells used. Thus, this novel system will be useful for SCNT-mediated acquisition of GM cloned piglets, in which multiple target loci may be mutated. | |||||
資源タイプ(コンテンツの種類) | ||||||
内容記述タイプ | Other | |||||
内容記述 | Article | |||||
ISSN | ||||||
収録物識別子タイプ | EISSN | |||||
収録物識別子 | 1422-0067 | |||||
書誌レコードID | ||||||
収録物識別子タイプ | NCID | |||||
収録物識別子 | AA12038549 | |||||
PubMed | ||||||
識別子タイプ | PMID | |||||
関連識別子 | https://www.ncbi.nlm.nih.gov/pubmed/29207527 | |||||
関連名称 | 29207527 | |||||
権利 | ||||||
権利情報 | © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (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 | |||||
WoS | ||||||
表示名 | Web of Science | |||||
URL | http://gateway.isiknowledge.com/gateway/Gateway.cgi?&GWVersion=2&SrcAuth=ShinshuUniv&SrcApp=ShinshuUniv&DestLinkType=FullRecord&DestApp=WOS&KeyUT=000418896700114 |