| Item type |
文献 / Documents(1) |
| 公開日 |
2021-07-09 |
| 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/10.1111/rda.13106 |
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関連名称 |
10.1111/rda.13106 |
| 出版タイプ |
|
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出版タイプ |
AM |
|
出版タイプResource |
http://purl.org/coar/version/c_ab4af688f83e57aa |
| タイトル |
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|
タイトル |
Effects of voltage strength during electroporation on the development and quality of in vitro-produced porcine embryos |
| タイトル別表記 |
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その他のタイトル |
Effects of voltage strength during electropolation on the development and quality of in vitro-produced porcine embryos |
| タイトル別表記 |
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その他のタイトル |
Electroporation conditions and embryonic development |
| 著者 |
ニシオ, カツトシ
谷原, 史倫
Nguyen, Thanh-Van
クニハラ, トシキ
ニイ, マサヒロ
平田, 真樹
竹本, 龍也
音井, 威重
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| 抄録 |
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内容記述 |
This study was conducted to determine suitable conditions for an experimental method in which the CRISPR/Cas9 system is introduced into in vitro-produced porcine zygotes by electroporation. In the first experiment, when putative zygotes derived from in vitro fertilization (IVF) were electroporated by either unipolar or bipolar pulses, keeping the voltage, pulse duration, and pulse number fixed at 30 V/mm, 1 msec, and five repeats, respectively, the rate of blastocyst formation from zygotes electroporated by bipolar pulses decreased compared to zygotes electroporated by unipolar pulses. In the second experiment, the putative zygotes were electroporated by electroporation voltages ranging from 20 V/mm – 40 V/mm with five 1-msec unipolar pulses. The rate of cleavage and blastocyst formation of zygotes electroporated at 40 V/mm was significantly lower (p < 0.05) than that of zygotes electroporated at less than 30 V/mm. Moreover, the apoptotic nuclei indices of blastocysts derived from zygotes electroporated by voltages greater than 30 V/mm significantly increased compared with those from zygotes electroporated by voltages less than 25 V/mm (p < 0.05). When zygotes were electroporated with Cas9 mRNA and single-guide RNA (sgRNA) targeting site in the FGF10 exon 3, the proportions of blastocysts with targeted genomic sequences were 7.7% (2/26) and 3.6% (1/28) in the embryos derived from zygotes electroporated at 25 V/mm and 30 V/mm, respectively. Our results indicate that electroporation at 25 V/mm may be an acceptable condition for introducing Cas9 mRNA and sgRNA into pig IVF zygotes under which the viability of the embryos is not significantly affected. |
| キーワード |
|
|
主題 |
CRISPR/Cas9 |
| キーワード |
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主題 |
electroporation |
| キーワード |
|
|
主題 |
genome editing |
| キーワード |
|
|
主題 |
in vitro fertilization |
| キーワード |
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主題 |
pig |
| 書誌情報 |
en : Reproduction in Domestic Animals
巻 53,
号 2,
p. 313-318,
発行日 2017-11-14
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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 |
14390531 |
| 出版者 |
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出版者 |
Wiley-VCH |
| 権利情報 |
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権利情報 |
This is the peer reviewed version of the following article: Nishio, K, Tanihara, F, Nguyen, T-V, et al. Effects of voltage strength during electroporation on the development and quality of in vitro-produced porcine embryos. Reprod Dom Anim. 2018; 53: 313– 318., which has been published in final form at https://doi.org/10.1111/rda.13106. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. |
| EID |
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|
識別子 |
346496 |
| 言語 |
|
|
言語 |
eng |
rda_53_2_313.pdf (708 KB)
