| Item type |
文献 / Documents(1) |
| 公開日 |
2023-03-03 |
| アクセス権 |
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|
アクセス権 |
open access |
| 資源タイプ |
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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 |
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|
関連識別子 |
https://doi.org/10.14202/vetworld.2022.2210-2216 |
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関連名称 |
10.14202/vetworld.2022.2210-2216 |
| 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 |
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|
タイトル |
Multiple gene editing in porcine embryos using a combination of microinjection, electroporation, and transfection methods |
| 著者 |
Le, Quynh Anh
Wittayarat, Manita
Namula, Zhao
Lin, Qingyi
タケバヤシ, コウキ
平田, 真樹
谷原, 史倫
Do, Lanh Thi Kim
音井, 威重
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| 抄録 |
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内容記述 |
Background and Aim: Mosaicism – the presence of both wild-type and mutant alleles – is a serious problem for zygotic gene modification through gene editing using the Clustered regularly interspaced short palindromic repeats-Cas9 (CRISPR/ Cas9) system. Different delivery methods, such as microinjection (MI), electroporation (EP), and transfection (TF), can be used to transfer CRISPR/Cas9 components into porcine zygotes. This study aimed to develop a method that combines MI, EP, and TF to improve mutation efficiency mediated through the CRISPR/Cas9 system for a triple-gene knockout in pigs.
Materials and Methods: The study consisted of three groups: The MI group with three simultaneously microinjected guide RNAs (gRNAs) targeting α-1,3-galactosyltransferase (GGTA1), cytidine 32 monophosphate-N-acetylneuraminic acid hydroxylase (CMAH), and β-1,4-N-acetyl-galactosaminyltransferase 2 (B4GALNT2); the MI + EP group with two gRNAs targeting GGTA1 and B4GALNT2 genes delivered into zygotes through MI, followed by EP of gRNA targeting the CMAH 1 h later; and the MI + EP + TF group with MI of gRNA targeting GGTA1 gene into zygotes, followed by EP of gRNA targeting CMAH 1 h later, and then TF of gRNA targeting the B4GALNT2 gene into zona-free zygotes after another hour.
Results: The rate of blastocysts carrying mutations in one or two gene(s) was significantly higher in the MI + EP + TF group than in the MI group. However, the blastocyst formation rate of zygotes in the MI + EP + TF group was lower than that of the zygotes in the other treatment groups.
Conclusion: The combination of CRISPR/Cas9 delivery methods may improve the mutation efficiency of triple-gene edited porcine blastocysts. |
| キーワード |
|
|
主題 |
clustered regularly interspaced short palindromic repeats/Cas9 |
| キーワード |
|
|
主題 |
electroporation |
| キーワード |
|
|
主題 |
microinjection |
| キーワード |
|
|
主題 |
porcine zygotes |
| キーワード |
|
|
主題 |
transfection |
| 書誌情報 |
en : Veterinary World
巻 15,
号 9,
p. 2210-2216,
発行日 2022-09-16
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| 収録物ID |
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収録物識別子タイプ |
ISSN |
|
収録物識別子 |
22310916 |
| 収録物ID |
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|
収録物識別子タイプ |
ISSN |
|
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. |
| EID |
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|
識別子 |
392739 |
| 言語 |
|
|
言語 |
eng |
vetworld_15_9_2210.pdf (852 KB)
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