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Efficient gene editing of pig embryos by combining electroporation and lipofection

https://tokushima-u.repo.nii.ac.jp/records/2013144
https://tokushima-u.repo.nii.ac.jp/records/2013144
c04ba36d-9351-4aaf-96e2-a683e7d3ad23
名前 / ファイル ライセンス アクション
vetworld_17_11_2701.pdf vetworld_17_11_2701.pdf (750 KB)
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Item type 文献 / Documents(1)
公開日 2025-04-11
アクセス権
アクセス権 open access
アクセス権URI http://purl.org/coar/access_right/c_abf2
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/10.14202/vetworld.2024.2701-2707
関連名称 10.14202/vetworld.2024.2701-2707
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 Efficient gene editing of pig embryos by combining electroporation and lipofection
著者 Lin, Qingyi

× Lin, Qingyi

en Lin, Qingyi

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Torigoe, Nanaka

× Torigoe, Nanaka

en Torigoe, Nanaka

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Liu, Bin

× Liu, Bin

en Liu, Bin

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Nakayama, Yuichiro

× Nakayama, Yuichiro

en Nakayama, Yuichiro

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中井, 綾

× 中井, 綾

WEKO 918
徳島大学 教育研究者総覧 337579/profile-ja.html

ja 中井, 綾

ja-Kana ナカイ, アヤ

en Nakai, Aya

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Namula, Zhao

× Namula, Zhao

en Namula, Zhao

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長原, 恵

× 長原, 恵

WEKO 1349
徳島大学 教育研究者総覧 395177/profile-ja.html

ja 長原, 恵

ja-Kana ナガハラ, メグミ

en Nagahara, Megumi

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谷原, 史倫

× 谷原, 史倫

WEKO 747
e-Rad 90754680
徳島大学 教育研究者総覧 289665/profile-ja.html

ja 谷原, 史倫
ISNI

ja-Kana タニハラ, フミノリ

en Tanihara, Fuminori

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平田, 真樹

× 平田, 真樹

WEKO 891
徳島大学 教育研究者総覧 327914/profile-ja.html

ja 平田, 真樹

ja-Kana ヒラタ, マキ

en Hirata, Maki

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音井, 威重

× 音井, 威重

WEKO 772
徳島大学 教育研究者総覧 292974/profile-ja.html
e-Rad_Researcher 30311814

ja 音井, 威重

ja-Kana オトイ, タケシゲ

en Otoi, Takeshige

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抄録
内容記述 Background and Aim: Mosaicism, which is characterized by the presence of wild-type and more than one mutant allele, poses a serious problem in zygotic gene modification through the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 system. Therefore, we used pig embryos to compare the gene editing efficiencies achieved by combining electroporation and lipofection using different aminopeptidase N (APN)-targeting guide RNA (gRNA) sequences.
Materials and Methods: Six gRNAs (gRNA1–6) with different target sequences were designed to target APN. Zona pellucida (ZP)-intact zygotes collected 10 h after the start of in vitro fertilization (IVF) were electroporated with each gRNA to compare their gene editing efficiency. The gRNA sequences that achieved the lowest and highest mutation rates (gRNA4 and gRNA6, respectively) were selected for additional lipofection to assess gene editing efficiency following combined treatment. As ZP removal is essential for lipofection, ZP-free zygotes were electroporated with gRNA4 or gRNA6 10 h after IVF initiation, followed by lipofection with the same gRNAs 24 or 29 h after IVF initiation. The electroporated ZP-intact and ZP-free zygotes were used as controls.
Results: gRNA4 and gRNA6 exhibited the lowest and highest mutation rates, respectively. gRNA4-targeted ZP-free embryos subjected to additional lipofection 29 h after IVF initiation exhibited significantly higher total and biallelic mutation rates than ZP-intact embryos that received only electroporation. Additional lipofection of gRNA6-targeted embryos had no obvious effect on mutation rates.
Conclusion: Electroporation combined with lipofection using gRNAs with low mutation rates may improve gene editing efficiency in pig embryos. However, the effects may vary based on the timing of gene editing.
キーワード
主題 electroporation
キーワード
主題 guide RNA sequence
キーワード
主題 lipofection
キーワード
主題 pig embryo
書誌情報 en : Veterinary World

巻 17, 号 11, p. 2701-2707, 発行日 2024-11-30
収録物ID
収録物識別子タイプ EISSN
収録物識別子 22310916
収録物ID
収録物識別子タイプ PISSN
収録物識別子 09728988
出版者
出版者 Veterinary World
権利情報
権利情報 This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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.
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識別子 416050
言語
言語 eng
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