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Efficient generation of GGTA1-deficient pigs by electroporation of the CRISPR/Cas9 system into in vitro-fertilized zygotes
https://tokushima-u.repo.nii.ac.jp/records/2008804
https://tokushima-u.repo.nii.ac.jp/records/200880446a97403-5e7d-4f41-ab5e-09ef1b686a31
| 名前 / ファイル | ライセンス | アクション |
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bmcbiotech_20_40.pdf (3.5 MB)
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| Item type | 文献 / Documents(1) | |||||||||||||||||||||||||||||||||||||
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| 公開日 | 2021-06-11 | |||||||||||||||||||||||||||||||||||||
| 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/10.1186/s12896-020-00638-7 | |||||||||||||||||||||||||||||||||||||
| 関連名称 | 10.1186/s12896-020-00638-7 | |||||||||||||||||||||||||||||||||||||
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| 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 | Efficient generation of GGTA1-deficient pigs by electroporation of the CRISPR/Cas9 system into in vitro-fertilized zygotes | |||||||||||||||||||||||||||||||||||||
| 著者 |
谷原, 史倫
× 谷原, 史倫
WEKO
747
× 平田, 真樹× Nguyen, Nhien Thi
× サワモト, オサム
× キクチ, タケシ
× ドイ, マサコ
× 音井, 威重
WEKO
772
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| 抄録 | ||||||||||||||||||||||||||||||||||||||
| 内容記述 | Background: Xenoantigens are a major source of concern with regard to the success of interspecific xenografts. GGTA1 encodes α1,3-galactosyltransferase, which is essential for the biosynthesis of galactosyl-alpha 1,3-galactose, the major xenoantigen causing hyperacute rejection. GGTA1-modified pigs, therefore, are promising donors for pig-to-human xenotransplantation. In this study, we developed a method for the introduction of the CRISPR/Cas9 system into in vitro-fertilized porcine zygotes via electroporation to generate GGTA1-modified pigs. Results: We designed five guide RNAs (gRNAs) targeting distinct sites in GGTA1. After the introduction of the Cas9 protein with each gRNA via electroporation, the gene editing efficiency in blastocysts developed from zygotes was evaluated. The gRNA with the highest gene editing efficiency was used to generate GGTA1-edited pigs. Six piglets were delivered from two recipient gilts after the transfer of electroporated zygotes with the Cas9/gRNA complex. Deep sequencing analysis revealed that five out of six piglets carried a biallelic mutation in the targeted region of GGTA1, with no off-target events. Furthermore, staining with isolectin B4 confirmed deficient GGTA1 function in GGTA1 biallelic mutant piglets. Conclusions: We established GGTA1-modified pigs with high efficiency by introducing a CRISPR/Cas9 system into zygotes via electroporation. Multiple gene modifications, including knock-ins of human genes, in porcine zygotes via electroporation may further improve the application of the technique in pig-to-human xenotransplantation. |
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| キーワード | ||||||||||||||||||||||||||||||||||||||
| 主題 | GGTA1 | |||||||||||||||||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||||||||||||||||
| 主題 | CRISPR/Cas9 | |||||||||||||||||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||||||||||||||||
| 主題 | Electroporation | |||||||||||||||||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||||||||||||||||
| 主題 | In vitro fertilization | |||||||||||||||||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||||||||||||||||
| 主題 | Pig | |||||||||||||||||||||||||||||||||||||
| 書誌情報 |
en : BMC Biotechnology 巻 20, p. 40, 発行日 2020-08-18 |
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| 収録物ID | ||||||||||||||||||||||||||||||||||||||
| 収録物識別子タイプ | ISSN | |||||||||||||||||||||||||||||||||||||
| 収録物識別子 | 14726750 | |||||||||||||||||||||||||||||||||||||
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| 収録物識別子タイプ | NCID | |||||||||||||||||||||||||||||||||||||
| 収録物識別子 | AA12034730 | |||||||||||||||||||||||||||||||||||||
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| 出版者 | BioMed Central | |||||||||||||||||||||||||||||||||||||
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| 出版者 | Springer Nature | |||||||||||||||||||||||||||||||||||||
| 権利情報 | ||||||||||||||||||||||||||||||||||||||
| 権利情報 | This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. | |||||||||||||||||||||||||||||||||||||
| EID | ||||||||||||||||||||||||||||||||||||||
| 識別子 | 373087 | |||||||||||||||||||||||||||||||||||||
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| 言語 | eng | |||||||||||||||||||||||||||||||||||||
