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One-Step Generation of Multiple Gene-Edited Pigs by Electroporation of the CRISPR/Cas9 System into Zygotes to Reduce Xenoantigen Biosynthesis
https://tokushima-u.repo.nii.ac.jp/records/2009256
https://tokushima-u.repo.nii.ac.jp/records/2009256f1caf9f6-0faa-443d-b840-2c995575fed3
| 名前 / ファイル | ライセンス | アクション |
|---|---|---|
ijms_22_5_2249.pdf (2.57 MB)
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| Item type | 文献 / Documents(1) | |||||||||||||||||||||||||
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| 公開日 | 2021-10-20 | |||||||||||||||||||||||||
| アクセス権 | ||||||||||||||||||||||||||
| アクセス権 | open access | |||||||||||||||||||||||||
| 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 | |||||||||||||||||||||||||
| 出版社版DOI | ||||||||||||||||||||||||||
| 関連識別子 | https://doi.org/10.3390/ijms22052249 | |||||||||||||||||||||||||
| 関連名称 | 10.3390/ijms22052249 | |||||||||||||||||||||||||
| 出版タイプ | ||||||||||||||||||||||||||
| 出版タイプ | VoR | |||||||||||||||||||||||||
| 出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||||||||||||||||||||||
| タイトル | ||||||||||||||||||||||||||
| タイトル | One-Step Generation of Multiple Gene-Edited Pigs by Electroporation of the CRISPR/Cas9 System into Zygotes to Reduce Xenoantigen Biosynthesis | |||||||||||||||||||||||||
| 著者 |
谷原, 史倫
× 谷原, 史倫× 平田, 真樹× Nguyen, Nhien Thi
× サワモト, オサム
× キクチ, タケシ
× 音井, 威重
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| 内容記述 | Xenoantigens cause hyperacute rejection and limit the success of interspecific xenografts. Therefore, genes involved in xenoantigen biosynthesis, such as GGTA1, CMAH, and B4GALNT2, are key targets to improve the outcomes of xenotransplantation. In this study, we introduced a CRISPR/Cas9 system simultaneously targeting GGTA1, CMAH, and B4GALNT2 into in vitro-fertilized zygotes using electroporation for the one-step generation of multiple gene-edited pigs without xenoantigens. First, we optimized the combination of guide RNAs (gRNAs) targeting GGTA1 and CMAH with respect to gene editing efficiency in zygotes, and transferred electroporated embryos with the optimized gRNAs and Cas9 into recipient gilts. Next, we optimized the Cas9 protein concentration with respect to the gene editing efficiency when GGTA1, CMAH, and B4GALNT2 were targeted simultaneously, and generated gene-edited pigs using the optimized conditions. We achieved the one-step generation of GGTA1/CMAH double-edited pigs and GGTA1/CMAH/B4GALNT2 triple-edited pigs. Immunohistological analyses demonstrated the downregulation of xenoantigens; however, these multiple gene-edited pigs were genetic mosaics that failed to knock out some xenoantigens. Although mosaicism should be resolved, the electroporation technique could become a primary method for the one-step generation of multiple gene modifications in pigs aimed at improving pig-to-human xenotransplantation. | |||||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||||
| 主題 | CRISPR/Cas9 | |||||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||||
| 主題 | GGTA1 | |||||||||||||||||||||||||
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| 主題 | CMAH | |||||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||||
| 主題 | B4GALNT2 | |||||||||||||||||||||||||
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| 主題 | pig | |||||||||||||||||||||||||
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| 主題 | in vitro fertilization | |||||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||||
| 主題 | electroporation | |||||||||||||||||||||||||
| 書誌情報 |
en : International Journal of Molecular Sciences 巻 22, 号 5, p. 2249, 発行日 2021-02-24 |
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| 収録物識別子タイプ | ISSN | |||||||||||||||||||||||||
| 収録物識別子 | 14220067 | |||||||||||||||||||||||||
| 出版者 | ||||||||||||||||||||||||||
| 出版者 | MDPI | |||||||||||||||||||||||||
| 権利情報 | ||||||||||||||||||||||||||
| 権利情報 | This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). | |||||||||||||||||||||||||
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| 識別子 | 374164 | |||||||||||||||||||||||||
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| 言語 | eng | |||||||||||||||||||||||||
