| Item type |
学術雑誌論文 / Journal Article(1) |
| 公開日 |
2019-09-09 |
| タイトル |
|
|
タイトル |
Timing of CRISPR/Cas9-related mRNA microinjection after activation as an important factor affecting genome editing efficiency in porcine oocytes |
| 言語 |
|
|
言語 |
eng |
| DOI |
|
|
|
関連識別子 |
https://doi.org/10.1016/j.theriogenology.2017.11.030 |
|
|
関連名称 |
10.1016/j.theriogenology.2017.11.030 |
| キーワード |
|
|
主題 |
alpha-Gal epitope, alpha-1,3-Galactosyltransferase, CRISPR/Cas9, Microinjection, mRNA, Mosaicism, Biallelic KO, Indels, Isolectin BS-I-B-4 |
| 資源タイプ |
|
|
資源 |
http://purl.org/coar/resource_type/c_6501 |
|
タイプ |
journal article |
| 著者 |
Sato, Masahiro
Kosuke, Maeda
Koriyama, Miyu
Inada, Emi
Saitoh, Issei
Ohtsuka, Masato
Nakamura, Shingo
Sakurai, Takayuki
Watanabe, Satoshi
Miyoshi, Kazuchika
|
| 信州大学研究者総覧へのリンク |
|
|
氏名 |
Sakurai, Takayuki |
|
URL |
http://soar-rd.shinshu-u.ac.jp/profile/ja.jaLUuNkh.html |
| 出版者 |
|
|
出版者 |
ELSEVIER SCIENCE INC |
| 引用 |
|
|
内容記述 |
THERIOGENOLOGY.108:29-38(2018) |
| 書誌情報 |
THERIOGENOLOGY
巻 108,
p. 29-38,
発行日 2018-03-01
|
| 抄録 |
|
|
内容記述 |
Recently, successful one-step genome editing by microinjection of CRISPR/Cas9-related mRNA components into the porcine zygote has been described. Given the relatively long gestational period and the high cost of housing swine, the establishment of an effective microinjection-based porcine genome editing method is urgently required. Previously, we have attempted to disrupt a gene encoding alpha-1,3-galactosyltransferase (GGTA1), which synthesizes the alpha-Gal epitope, by microinjecting CRISPR/Cas9-related nucleic acids and enhanced green fluorescent protein (EGFP) mRNA into porcine oocytes immediately after electrical activation. We found that genome editing was indeed induced, although the resulting blastocysts were mosaic and the frequency of modified cells appeared to be low (50%). To improve genome editing efficiency in porcine oocytes, cytoplasmic injection was performed 6 h after electrical activation, a stage wherein the pronucleus is formed. The developing blastocysts exhibited higher levels of EGFP. Furthermore, the T7 endonuclease 1 assay and subsequent sequencing demonstrated that these embryos exhibited increased genome editing efficiencies (69%), although a high degree of mosaicism for the induced mutation was still observed. Single blastocyst-based cytochemical staining with fluorescently labeled isolectin BS-I-B-4 also confirmed this mosaicism. Thus, the development of a technique that avoids or reduces such mosaicism would be a key factor for efficient knock out piglet production via microinjection. (C) 2017 Elsevier Inc. All rights reserved. |
| 資源タイプ（コンテンツの種類） |
|
| ISSN |
|
|
収録物識別子タイプ |
PISSN |
|
収録物識別子 |
0093-691X |
| 書誌レコードID |
|
|
収録物識別子タイプ |
NCID |
|
収録物識別子 |
AA00862972 |
| PubMed |
|
|
|
識別子タイプ |
PMID |
|
|
関連識別子 |
https://www.ncbi.nlm.nih.gov/pubmed/29195121 |
|
|
関連名称 |
29195121 |
| 権利 |
|
|
権利情報 |
© 2018. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/ |
| 出版タイプ |
|
|
出版タイプ |
AM |
|
出版タイプResource |
http://purl.org/coar/version/c_ab4af688f83e57aa |
| WoS |
|
|
URL |
http://gateway.isiknowledge.com/gateway/Gateway.cgi?&GWVersion=2&SrcAuth=ShinshuUniv&SrcApp=ShinshuUniv&DestLinkType=FullRecord&DestApp=WOS&KeyUT=000424732900005 |
16K15233_01.pdf (3.1 MB)
