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Establishment of protocol for preparation of gene-edited bovine ear-derived fibroblasts for somatic cell nuclear transplantation
http://hdl.handle.net/10659/00006308
http://hdl.handle.net/10659/000063086f9ed494-9771-4733-b29d-b57b4735a6f9
名前 / ファイル | ライセンス | アクション |
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R-2019-44_kitamura.pdf (1.7 MB)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2020-02-26 | |||||
タイトル | ||||||
タイトル | Establishment of protocol for preparation of gene-edited bovine ear-derived fibroblasts for somatic cell nuclear transplantation | |||||
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 | |||||
著者 |
ISHINO, Takeshi
× ISHINO, Takeshi× HASHIMOTO, Mayuko× AMAGASA, Misato× SAITO, Natsuko× 堂地, 修× KIRISAWA, Rikio× 北村, 浩 |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Recently, gene-editing using the clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated protein 9 (Cas9) technique has attempted to utilize fibroblasts of livestock animals for somatic cell nuclear transfer. In this study, we establish the procedure for preparing skin fibroblast clones whose genes were edited by the CRISPR/Cas9 technique. After isolating fibroblasts from earlobes of Japanese Black cattle, subsequent collagenase-digestion and extensive wash procedures enabled us to avoid contamination of fungi. Electroporation using NEPA21, rather than lipofection using commercially available liposome reagents, allowed us to perform more efficient transfection of plasmid constructs. Although bovine ear-derived fibroblasts were not able to proliferate in single cell cultures in Dulbecco’s modified Eagle medium containing 10% fetal calf serum, supplementation with insulin-transferrin-selenium mixture, human recombinant epidermal growth factor, or human recombinant basic fibroblast growth factor promoted proliferation of the cells, even in a single cell culture. Taking advantage of our established protocol, we eventually obtained eight ear-derived fibroblast clones with a recessive mutation in the isoleucyl-tRNA synthetase gene corrected by the CRISPR/Cas9 technique. | |||||
書誌情報 |
Biomedical Research 巻 39, 号 2, p. 95-104, 発行日 2018 |
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ISBN | ||||||
識別子タイプ | ISBN | |||||
関連識別子 | 0388-6107 | |||||
DOI | ||||||
関連タイプ | isIdenticalTo | |||||
識別子タイプ | DOI | |||||
関連識別子 | http://dx.doi.org/10.2220/biomedres.39.95 | |||||
権利 | ||||||
権利情報 | © 2018 Biomedical Research Press | |||||
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 | |||||
出版者 | ||||||
出版者 | Biomedical Research Press | |||||
資源タイプ | ||||||
内容記述タイプ | Other | |||||
内容記述 | Article |