| Item type |
文献 / Documents(1) |
| 公開日 |
2022-04-19 |
| アクセス権 |
|
|
アクセス権 |
open access |
| 資源タイプ |
|
|
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/10.1186/s13104-021-05800-8 |
|
|
関連名称 |
10.1186/s13104-021-05800-8 |
| 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 |
Timing and duration of lipofection-mediated CRISPR/Cas9 delivery into porcine zygotes affect gene-editing events |
| 著者 |
Lin, Qingyi
Le, Quynh Anh
タケバヤシ, コウキ
ソンキテイデイロ, チヨンマナート
| ja |
ソンキテイデイロ, チヨンマナート
|
| ja-Kana |
トンキッチィディロク, チヨンマナート
|
| en |
Thongkittidilok, Chommanart
|
Search repository
Wittayarat, Manita
平田, 真樹
谷原, 史倫
音井, 威重
|
| 抄録 |
|
|
内容記述 |
Objective: Lipofection-mediated introduction of the CRISPR/Cas9 system in porcine zygotes provides a simple method for gene editing, without requiring micromanipulation. However, the gene editing efficiency is inadequate. The aim of this study was to improve the lipofection-mediated gene editing efficiency by optimizing the timing and duration of lipofection.
Results: Zona pellucida (ZP)-free zygotes collected at 5, 10, and 15 h from the start of in vitro fertilization (IVF) were incubated with lipofection reagent, guide RNA (gRNA) targeting GGTA1, and Cas9 for 5 h. Lipofection of zygotes collected at 10 and 15 h from the start of IVF yielded mutant blastocysts. Next, ZP-free zygotes collected at 10 h from the start of IVF were incubated with lipofection reagent, gRNA, and Cas9 for 2.5, 5, 10, or 20 h. The blastocyst formation rate of zygotes treated for 20 h was significantly lower (p < 0.05) than those of the other groups, and no mutant blastocysts were obtained. Moreover, the mutation rates of the resulting blastocysts decreased as the incubation time increased. In conclusion, a lipofection-mediated gene editing system using the CRISPR/Cas9 system in ZP-zygotes is feasible; however, further improvements in the gene editing efficiency are required. |
| キーワード |
|
|
主題 |
Lipofection |
| キーワード |
|
|
主題 |
CRISPR |
| キーワード |
|
|
主題 |
Cas9 |
| キーワード |
|
|
主題 |
In vitro fertilized zygote |
| キーワード |
|
|
主題 |
Pig |
| 書誌情報 |
en : BMC Research Notes
巻 14,
p. 389,
発行日 2021-10-09
|
| 収録物ID |
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|
収録物識別子タイプ |
ISSN |
|
収録物識別子 |
17560500 |
| 出版者 |
|
|
出版者 |
BioMed Central |
| 出版者 |
|
|
出版者 |
Springer Nature |
| 権利情報 |
|
|
権利情報 |
This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. 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 in a credit line to the data. |
| EID |
|
|
識別子 |
384280 |
| 言語 |
|
|
言語 |
eng |
BMCRN_14_389.pdf (744 KB)
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