| Item type |
文献 / Documents(1) |
| 公開日 |
2025-02-26 |
| アクセス権 |
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|
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 |
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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 |
|
|
|
関連識別子 |
https://doi.org/10.1262/jrd.2024-054 |
|
|
関連名称 |
10.1262/jrd.2024-054 |
| 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 |
Genome editing of porcine zygotes via lipofection of two guide RNAs using a CRISPR/Cas9 system |
| タイトル別表記 |
|
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その他のタイトル |
DOUBLE GENE EDITING USING LIPOFECTION |
| 著者 |
Lin, Qingyi
Takebayashi, Koki
Torigoe, Nanaka
Liu, Bin
Namula, Zhao
平田, 真樹
谷原, 史倫
長原, 恵
音井, 威重
|
| 抄録 |
|
|
内容記述 |
CRISPR/Cas9-based multiplex genome editing via electroporation is relatively efficient; however, lipofection is versatile because of its ease of use and low cost. Here, we aimed to determine the efficiency of lipofection in CRISPR/Cas9-based multiplex genome editing using growth hormone receptor (GHR) and glycoprotein alpha-galactosyltransferase 1 (GGTA1)-targeting guide RNAs (gRNAs) in pig zygotes. Zona pellucida-free zygotes were collected 10 h after in vitro fertilization and incubated with Cas9, gRNAs, and Lipofectamine 2000 (LP2000) for 5 h. In Experiment 1, we evaluated the mutation efficiency of gRNAs targeting either GHR or GGTA1 in zygotes transfected using LP2000 and cultured in 4-well plates. In Experiment 2, we examined the effects of the culture method on the development, mutation rate, and mutation efficiency of zygotes with simultaneously double-edited GHR and GGTA1, cultured using 4-well (group culture) and 25-well plates (individual culture). In Experiment 3, we assessed the effect of additional GHR-targeted lipofection before and after simultaneous double gRNA-targeted lipofection on the mutation efficiency of edited embryos cultured in 25-well plates. No significant differences in mutation rates were observed between the zygotes edited with either gRNA. Moreover, the formation rate of blastocysts derived from GHR and GGTA1 double-edited zygotes was significantly increased in the 25-well plate culture compared to that in the 4-well plate culture. However, mutations were only observed in GGTA1 when zygotes were transfected with both gRNAs, irrespective of the culture method used. GHR mutations were detected only in blastocysts derived from zygotes subjected to GHR-targeted lipofection before simultaneous double gRNA-targeted lipofection. Overall, our results suggest that additional lipofection before simultaneous double gRNA-targeted lipofection induces additional mutations in the zygotes. |
| キーワード |
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|
主題 |
CRISPR/Cas9 system |
| キーワード |
|
|
主題 |
Growth hormone receptor (GHR) |
| キーワード |
|
|
主題 |
Glycoprotein alpha-galactosyltransferase 1 (GGTA1) |
| キーワード |
|
|
主題 |
Lipofection |
| キーワード |
|
|
主題 |
Porcine zygote |
| 書誌情報 |
en : Journal of Reproduction and Development
巻 70,
号 6,
p. 356-361,
発行日 2024
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| 収録物ID |
|
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収録物識別子タイプ |
EISSN |
|
収録物識別子 |
13484400 |
| 収録物ID |
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|
収録物識別子タイプ |
PISSN |
|
収録物識別子 |
09168818 |
| 出版者 |
|
|
出版者 |
The Society for Reproduction and Development |
| 権利情報 |
|
|
権利情報 |
This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. (CC-BY-NC-ND 4.0: https://creativecommons.org/licenses/by-nc-nd/4.0/) |
| EID |
|
|
識別子 |
414735 |
| 言語 |
|
|
言語 |
eng |
jrd_70_6_356.pdf (1.2 MB)
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