| Item type |
文献 / Documents(1) |
| 公開日 |
2023-01-18 |
| アクセス権 |
|
|
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/10.1016/j.vas.2022.100241 |
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関連名称 |
10.1016/j.vas.2022.100241 |
| 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 |
Effects of the timing of electroporation during in vitro maturation on triple gene editing in porcine embryos using CRISPR/Cas9 system |
| 著者 |
Namula, Zhao
Wittayarat, Manita
Do, Lanh Thi Kim
Nguyen, Thanh Van
Lin, Qingyi
タケバヤシ, コウキ
平田, 真樹
谷原, 史倫
音井, 威重
|
| 抄録 |
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内容記述 |
Mosaicism, including alleles comprising both wild-type and mutant, is a serious problem for gene modification by gene editing using electroporation. One-step generation of F0 pigs with completely desired gene modifications saves cost and time, but the major obstacles have been mosaic mutations. We hypothesized that the timing of electroporation prior to in vitro fertilization (IVF) can increase the rates of biallelic mutation for multiple gene knockout as the permeability of mature oocytes is greater than that of zygotes. Hence, we determined whether the timing of electroporation during in vitro maturation (IVM) culture enhances triple gene editing in the resulting blastocysts. Three gRNAs targeting KDR, PDX1, and SALL1 were simultaneously introduced into the oocytes that had been incubated for 40, 42, and 44 h from the start of the IVM culture. Electroporation with three gRNAs at 40 h and 42 h during IVM culture decreased the blastocyst formation rates and did not improve the mutation rates and target number of biallelic mutations in the resulting blastocysts. The blastocyst formation rate, mutation rates, and target numbers in the resulting blastocysts from oocytes treated by electroporation at 44 h of IVM culture were similar to those of control zygotes electroporated at 13 h after the initiation of IVF. In conclusion, multiple gene editing efficiency in the resulting blastocysts was comparable between oocytes electroporated before and after the fertilization, indicating that oocytes with completed maturation time may allow better functioning of materials accepting gene editing application. |
| キーワード |
|
|
主題 |
CRISPR/Cas9 |
| キーワード |
|
|
主題 |
Electroporation |
| キーワード |
|
|
主題 |
Gene knockout |
| キーワード |
|
|
主題 |
Permeability |
| キーワード |
|
|
主題 |
Pig |
| 書誌情報 |
en : Veterinary and Animal Science
巻 16,
p. 100241,
発行日 2022-02-28
|
| 収録物ID |
|
|
収録物識別子タイプ |
ISSN |
|
収録物識別子 |
2451943X |
| 出版者 |
|
|
出版者 |
Elsevier |
| 権利情報 |
|
|
権利情報 |
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
| EID |
|
|
識別子 |
387619 |
| 言語 |
|
|
言語 |
eng |
vas_16_100241.pdf (578 KB)
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