| Item type |
文献 / Documents(1) |
| 公開日 |
2021-02-19 |
| 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 |
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資源タイプ |
journal article |
| 出版社版DOI |
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関連識別子 |
https://doi.org/10.1262/jrd.2018-116 |
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関連名称 |
10.1262/jrd.2018-116 |
| 出版タイプ |
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出版タイプ |
VoR |
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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 |
Effects of concentration of CRISPR/Cas9 components on genetic mosaicism in cytoplasmic microinjected porcine embryos |
| タイトル別表記 |
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その他のタイトル |
EMBRYONIC MOSAICISM BY MICROINJECTION |
| 著者 |
谷原, 史倫
平田, 真樹
Nguyen, Nhien Thi
Le, Quynh Anh
ヒラノ, タカユキ
音井, 威重
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| 抄録 |
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内容記述 |
Cytoplasmic microinjection (CI) of the CRISPR/Cas9 system enabled the induction of site-specific mutations in porcine zygotes and resulting pigs. However, mosaicism is a serious problem for genetically modified pigs. In the present study, we investigated suitable timing and concentration of CRISPR/Cas9 components for introduction into oocytes/zygotes by CI, to reduce mosaicism in the resulting blastocysts. First, we introduced 20 ng/μl of Cas9 protein and guide RNA (gRNA), targeting the α-1,3-galactosyltransferase (GalT) gene in oocytes before in vitro fertilization (IVF), in zygotes after IVF, or in oocytes/zygotes before and after IVF, twice. CI treatment had no detrimental effects on blastocyst formation rates. The highest value of the rate of mutant blastocysts was observed in zygotes injected after IVF. Next, we injected Cas9 protein and gRNA into zygotes after IVF at a concentration of 20 ng/μl each (20 ng/μl group) or 100 ng/μl each (100 ng/μl group). The ratio of the number of blastocysts that carried mutations to the total number of blastocysts examined in the 100 ng/μl group was significantly higher (P < 0.05) than that in the 20 ng/μl group. Although no blastocysts from the 20 ng/μl group carried a biallelic mutation, 16.7% of blastocysts from the 100 ng/μl group carried a biallelic mutation. In conclusion, increasing the concentration of Cas9 protein and gRNA is effective in generating biallelic mutant blastocysts. To reduce mosaicism, however, further optimization of the timing of CI, and the concentration of CRISPR/Cas9 components, is needed. |
| キーワード |
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主題 |
CRISPR/Cas9 |
| キーワード |
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|
主題 |
Cytoplasmic microinjection |
| キーワード |
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|
主題 |
Gene editing |
| キーワード |
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|
主題 |
Mosaicism |
| キーワード |
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主題 |
Porcine zygote |
| 書誌情報 |
en : Journal of Reproduction and Development
巻 65,
号 3,
p. 209-214,
発行日 2019
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| 収録物ID |
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収録物識別子タイプ |
ISSN |
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収録物識別子 |
13484400 |
| 収録物ID |
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収録物識別子タイプ |
ISSN |
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収録物識別子 |
09168818 |
| 収録物ID |
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収録物識別子タイプ |
NCID |
|
収録物識別子 |
AA10936678 |
| 出版者 |
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出版者 |
Society for Reproduction and Development |
| 権利情報 |
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|
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 |
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識別子 |
350607 |
| 言語 |
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言語 |
eng |
jrd_65_3_209.pdf (966 KB)
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