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Lipofection-Mediated Introduction of CRISPR/Cas9 System into Porcine Oocytes and Embryos
https://tokushima-u.repo.nii.ac.jp/records/2009190
https://tokushima-u.repo.nii.ac.jp/records/20091908299fa0b-958f-4325-a1ff-76b0995d12b9
| 名前 / ファイル | ライセンス | アクション |
|---|---|---|
ani_11_2_578.pdf (782 KB)
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| Item type | 文献 / Documents(1) | |||||||||||||||||||||||||||||||
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| 公開日 | 2021-09-13 | |||||||||||||||||||||||||||||||
| 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/ani11020578 | |||||||||||||||||||||||||||||||
| 関連名称 | 10.3390/ani11020578 | |||||||||||||||||||||||||||||||
| 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 | Lipofection-Mediated Introduction of CRISPR/Cas9 System into Porcine Oocytes and Embryos | |||||||||||||||||||||||||||||||
| 著者 |
平田, 真樹
× 平田, 真樹× Wittayarat, Manita
× Namula, Zhao
× Le, Quynh Anh
× Lin, Qingyi
× タケバヤシ, コウキ
× ソンキテイデイロ, チヨンマナート
× 谷原, 史倫× 音井, 威重
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| 内容記述 | Liposome-mediated gene transfer has become an alternative method for establishing a gene targeting framework, and the production of mutant animals may be feasible even in laboratories without specialized equipment. However, whether blastocyst genome editing can be performed by treatment with lipofection reagent, guide RNA, and Cas9, without performing electroporation or microinjection, remains unclear. In this study, we demonstrated that lipofection treatment successfully induced mutation into zygotes during in vitro fertilization and in embryos at the 2- and 4-cell stages. Although liposome-mediated gene editing is a feasible system for use with zona-pellucida-free oocytes/embryos, several challenges must be overcome. | |||||||||||||||||||||||||||||||
| 抄録 | ||||||||||||||||||||||||||||||||
| 内容記述 | Liposome-mediated gene transfer has become an alternative method for establishing a gene targeting framework, and the production of mutant animals may be feasible even in laboratories without specialized equipment. However, how this system functions in mammalian oocytes and embryos remains unclear. The present study was conducted to clarify whether blastocyst genome editing can be performed by treatment with lipofection reagent, guide RNA, and Cas9 for 5 h without using electroporation or microinjection. A mosaic mutation was observed in blastocysts derived from zona pellucida (ZP)-free oocytes following lipofection treatment, regardless of the target genes. When lipofection treatment was performed after in vitro fertilization (IVF), no significant differences in the mutation rates or mutation efficiency were found between blastocysts derived from embryos treated at 24 and 29 h from the start of IVF. Only blastocysts from embryos exposed to lipofection treatment at 29 h after IVF contained biallelic mutant. Furthermore, there were no significant differences in the mutation rates or mutation efficiency between blastocysts derived from embryos at the 2- and 4-cell stages. This suggests that lipofection-mediated gene editing can be performed in ZP-free oocytes and ZP-free embryos; however, other factors affecting the system efficiency should be further investigated. | |||||||||||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||||||||||
| 主題 | CRISPR/Cas9 | |||||||||||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||||||||||
| 主題 | embryo | |||||||||||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||||||||||
| 主題 | lipofection | |||||||||||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||||||||||
| 主題 | oocyte | |||||||||||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||||||||||
| 主題 | pig | |||||||||||||||||||||||||||||||
| 書誌情報 |
en : Animals 巻 11, 号 2, p. 578, 発行日 2021-02-23 |
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| 収録物識別子タイプ | ISSN | |||||||||||||||||||||||||||||||
| 収録物識別子 | 20762615 | |||||||||||||||||||||||||||||||
| 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/). | |||||||||||||||||||||||||||||||
| EID | ||||||||||||||||||||||||||||||||
| 識別子 | 374163 | |||||||||||||||||||||||||||||||
| 言語 | ||||||||||||||||||||||||||||||||
| 言語 | eng | |||||||||||||||||||||||||||||||
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平田, 真樹, Wittayarat, Manita, Namula, Zhao, Le, Quynh Anh, Lin, Qingyi, タケバヤシ, コウキ, ソンキテイデイロ, チヨンマナート, 谷原, 史倫, 音井, 威重, 2021, Lipofection-Mediated Introduction of CRISPR/Cas9 System into Porcine Oocytes and Embryos: MDPI, 578– p.
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