ArticleIn vitro cellular & developmental biology. Animal2019
Genome mutation after introduction of the gene editing by electroporation of Cas9 protein (GEEP) system in matured oocytes and putative zygotes.
Article in In vitro cellular & developmental biology. Animal, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 33 citations in OpenAlex.
- Double Electroporation Combined with Zona Pellucida Removal Improves Biallelic Genome Editing Efficiency in Porcine Embryos.Animals : an open access journal from MDPI · 2026Article
- Effects of centrifugation treatment before electroporation on gene editing in pig embryos.In vitro cellular & developmental biology. Animal · 2024Article
- Effects of the timing of electroporation duringVeterinary and animal science · 2022Article
- Loss of heterochromatin and retrotransposon silencing as determinants in oocyte aging.Aging cell · 2022Article
- Effect of Aphidicolin, a Reversible Inhibitor of Eukaryotic Nuclear DNA Replication, on the Production of Genetically Modified Porcine Embryos by CRISPR/Cas9.International journal of molecular sciences · 2022Article
- Current status of the application of gene editing in pigs.The Journal of reproduction and development · 2021Review
- Electroporation-Mediated Genome Editing of Livestock Zygotes.Frontiers in genetics · 2021Review
- One-step genome editing of porcine zygotes through the electroporation of a CRISPR/Cas9 system with two guide RNAs.In vitro cellular & developmental biology. Animal · 2020Article
- Efficient generation of GGTA1-deficient pigs by electroporation of the CRISPR/Cas9 system into in vitro-fertilized zygotes.BMC biotechnology · 2020Article
- CRISPR in livestock: From editing to printing.Theriogenology · 2020Review
- Efficient One-Step Knockout by Electroporation of Ribonucleoproteins Into Zona-Intact Bovine Embryos.Frontiers in genetics · 2020Article
- Genome mutation after the introduction of the gene editing by electroporation of Cas9 protein (GEEP) system into bovine putative zygotes.In vitro cellular & developmental biology. Animal · 2019Article
Corrections and comments
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Authors and funding
9 authors at 4 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The application of CRISPR/Cas9 strategy promises to rapidly increase the production of genetically engineered animals since it yields stably integrated transgenes. In the present study, we investigated the efficiency of target mutations after electroporation with the CRISPR/Cas9 system using sgRNAs to target the MSTN or FGF10 genes in porcine-matured oocytes and putative zygotes. Effects of pulse number (3-7 pulse repetitions) during electroporation on the embryonic development and mutation efficiency were also investigated. Our results showed that the cleavage rate of matured oocytes with electroporation treatment significantly decreased as compared with electroporated putative zygotes (p < 0.05). Moreover, the rates of blastocyst formation from oocytes/zygotes electroporated with more than 5 pulses decreased. Mutation efficiency was then assessed after sequencing the target sites in individual blastocysts derived from oocytes/zygotes electroporated by 3 and 5 pulses. No bi-allelic mutations in all examined blastocysts were observed in this study. There were no differences in the mutation rates (50-60%) between blastocysts derived from matured oocytes electroporated by 3 and 5 pulses, irrespective of targeting gene. In the targeting MSTN gene, however, the mutation rate (12.5%) of blastocysts derived from putative zygotes electroporated by 3 pulses tended to be lower than that (60%) from 5-pulsed electroporated putative zygotes. These data indicate that the type of eggs may influence not only their development after electroporation treatment but also the mutation rate in the resulting blastocysts.
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