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  1. 050 医学部, 大学院医学系研究科
  2. 0501 学術論文

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
5038cfa4-3c6f-4b85-8a7e-636eb806f017
名前 / ファイル ライセンス アクション
16K15233_04.pdf 16K15233_04.pdf (25.5 MB)
Item type 学術雑誌論文 / Journal Article(1)
公開日 2019-09-09
タイトル
言語 en
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
言語 eng
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 http://purl.org/coar/resource_type/c_6501
タイプ journal article
著者 Sato, Masahiro

× Sato, Masahiro

WEKO 108329

en Sato, Masahiro

Search repository
Miyoshi, Kazuchika

× Miyoshi, Kazuchika

WEKO 108330

en Miyoshi, Kazuchika

Search repository
Nakamura, Shingo

× Nakamura, Shingo

WEKO 108331

en Nakamura, Shingo

Search repository
Ohtsuka, Masato

× Ohtsuka, Masato

WEKO 108332

en Ohtsuka, Masato

Search repository
Sakurai, Takayuki

× Sakurai, Takayuki

WEKO 108333

en Sakurai, Takayuki

Search repository
Watanabe, Satoshi

× Watanabe, Satoshi

WEKO 108334

en Watanabe, Satoshi

Search repository
Kawaguchi, Hiroaki

× Kawaguchi, Hiroaki

WEKO 108335

en Kawaguchi, Hiroaki

Search repository
Tanimoto, Akihide

× Tanimoto, Akihide

WEKO 108336

en Tanimoto, Akihide

Search repository
信州大学研究者総覧へのリンク
氏名 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
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
関連識別子
識別子タイプ PMID
関連識別子 https://www.ncbi.nlm.nih.gov/pubmed/29207527
関連名称
関連名称 29207527
DOI
関連識別子
識別子タイプ DOI
関連識別子 https://doi.org/10.3390/ijms18122610
関連名称
関連名称 10.3390/ijms18122610
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
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