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ID 116985
Author
Namula, Zhao Guangdong Ocean University|Tokushima University
Le, Quynh Anh Tokushima University
Lin, Qingyi Tokushima University
Takebayashi, Koki Tokushima University
Yoshimura, Naoaki Tokushima University
Thongkittidilok, Chommanart Walailak University
Keywords
CRISPR
electroporation
genome editing
porcine zygote
zona pellucida
Content Type
Journal Article
Description
Background: Increasing the permeability of the zona pellucida (ZP) of oocytes before CRISPR/Cas9 electroporation may improve the efficiency of gene editing; however, the effects of this approach on subsequent developmental processes are unclear. In this study, the effects of ZP treatment before electroporation on embryonic development and gene editing in porcine embryos were evaluated.
Methods: The ZP of zygotes was weakened or removed by exposure to 0.5% actinase E, followed by electroporation of the Cas9 protein with guide RNA targeting GGTA1.
Results: The blastocyst formation rate of ZP-free zygotes after electroporation was significantly lower (p < 0.05) than that of ZP-intact zygotes. The mutation rate in blastocysts from ZP-weakened zygotes was similar to that in ZP-intact zygotes, whereas ZP removal increased the mutation rate. The mutation efficiency in blastocysts from electroporated zygotes did not differ among ZP treatment groups.
Conclusions: Our results indicate that weakening the ZP does not affect the developmental competence, mutation rate, or mutation efficiency of electroporated zygotes, whereas ZP removal has a detrimental effect on embryonic development but may increase the mutation rate.
Journal Title
Veterinary Medicine and Science
ISSN
20531095
Publisher
John Wiley & Sons
Volume
8
Issue
1
Start Page
164
End Page
169
Published Date
2021-10-21
Rights
This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/), which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
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DOI (Published Version)
URL ( Publisher's Version )
FullText File
language
eng
TextVersion
Publisher
departments
Bioscience and Bioindustry