ArticleInternational journal of molecular sciences2015
Direct Injection of CRISPR/Cas9-Related mRNA into Cytoplasm of Parthenogenetically Activated Porcine Oocytes Causes Frequent Mosaicism for Indel Mutations.
Article in International journal of molecular sciences, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed, 74 citations in OpenAlex.
- Development of a Genetically Engineered Porcine Model of Rhabdoid Tumor Predisposition Syndrome Type 1 (RTPS-1).Cancers · 2026Article
- A bibliometric mapping of advancements and trends in genome editing in pigs.Tropical animal health and production · 2025Review
- AeLife · 2024Article
- Fetal bone engraftment reconstitutes the immune system in pigs with severe combined immunodeficiency.Lab animal · 2024Article
- Evaluation of guide-free Cas9-induced genomic damage and transcriptome changes in pig embryos.Molecular therapy. Nucleic acids · 2023Article
- CRISPR-Cas9-Based Technology and Its Relevance to Gene Editing in Parkinson's Disease.Pharmaceutics · 2022Review
- Impact of CRISPR-Cas9-Based Genome Engineering in Farm Animals.Veterinary sciences · 2021Review
- Porcine germline genome engineering.Proceedings of the National Academy of Sciences of the United States of America · 2021Review
- CRISPR/Cas9 Mediated High Efficiency Knockout of Myosin Essential Light Chain Gene in the Pacific Oyster (Crassostrea Gigas).Marine biotechnology (New York, N.Y.) · 2021Article
- Comparison of the effects of introducing the CRISPR/Cas9 system by microinjection and electroporation into porcine embryos at different stages.BMC research notes · 2021Article
- Electroporation-Mediated Genome Editing of Livestock Zygotes.Frontiers in genetics · 2021Review
- Hydrodynamics-Based Transplacental Delivery as a Useful Noninvasive Tool for Manipulating Fetal Genome.Cells · 2020Article
- Sequential i-GONAD: An Improved In Vivo Technique for CRISPR/Cas9-Based Genetic Manipulations in Mice.Cells · 2020Article
- Suppression of mosaic mutation by co-delivery of CRISPR associated protein 9 and three-prime repair exonuclease 2 into porcine zygotes via electroporation.The Journal of reproduction and development · 2020Article
- Production of genetically engineered mice with higher efficiency, lower mosaicism, and multiplexing capability using maternally expressed Cas9.Scientific reports · 2020Article
- Practical Approaches for Knock-Out Gene Editing in Pigs.Frontiers in genetics · 2020Review
- Efficient One-Step Knockout by Electroporation of Ribonucleoproteins Into Zona-Intact Bovine Embryos.Frontiers in genetics · 2020Article
- Strategies to reduce genetic mosaicism following CRISPR-mediated genome edition in bovine embryos.Scientific reports · 2019Article
- Human germline editing: Insights to future clinical treatment of diseases.Protein & cell · 2019Review
- Effects of concentration of CRISPR/Cas9 components on genetic mosaicism in cytoplasmic microinjected porcine embryos.The Journal of reproduction and development · 2019Article
Corrections and comments
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Authors and funding
9 authors at 6 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Some reports demonstrated successful genome editing in pigs by one-step zygote microinjection of mRNA of CRISPR/Cas9-related components. Given the relatively long gestation periods and the high cost of housing, the establishment of a single blastocyst-based assay for rapid optimization of the above system is required. As a proof-of-concept, we attempted to disrupt a gene (GGTA1) encoding the α-1,3-galactosyltransferase that synthesizes the α-Gal epitope using parthenogenetically activated porcine oocytes. The lack of α-Gal epitope expression can be monitored by staining with fluorescently labeled isolectin BS-I-B4 (IB4), which binds specifically to the α-Gal epitope. When oocytes were injected with guide RNA specific to GGTA1 together with enhanced green fluorescent protein (EGFP) and human Cas9 mRNAs, 65% (24/37) of the developing blastocysts exhibited green fluorescence, although almost all (96%, 23/24) showed a mosaic fluorescent pattern. Staining with IB4 revealed that the green fluorescent area often had a reduced binding activity to IB4. Of the 16 samples tested, six (five fluorescent and one non-fluorescent blastocysts) had indel mutations, suggesting a correlation between EGFP expression and mutation induction. Furthermore, it is suggested that zygote microinjection of mRNAs might lead to the production of piglets with cells harboring various mutation types.
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