ArticleBMC biotechnology2020
Efficient generation of GGTA1-deficient pigs by electroporation of the CRISPR/Cas9 system into in vitro-fertilized zygotes.
Article in BMC biotechnology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 41 citations in OpenAlex.
- Advances in pig embryo production for cryopreservation, transfer and genome editing.The Journal of reproduction and development · 2026Review
- Comparison Between Electroporation at Different Voltage Levels and Microinjection to Generate Porcine Embryos with Multiple Xenoantigen Knock-Outs.International journal of molecular sciences · 2024Article
- CRISPR Technology Acts as a Dual-Purpose Tool in Pig Breeding: Enhancing Both Agricultural Productivity and Biomedical Applications.Biomolecules · 2024Review
- Programmed cell death-1-modified pig developed using electroporation-mediated gene editing for in vitro fertilized zygotes.In vitro cellular & developmental biology. Animal · 2024Article
- GHR-mutant pig derived from domestic pig and microminipig hybrid zygotes using CRISPR/Cas9 system.Molecular biology reports · 2023Article
- - Invited Review - Reproductive technologies needed for the generation of precise gene-edited pigs in the pathways from laboratory to farm.Animal bioscience · 2023Article
- Electroporation-Mediated CRISPR/Cas9 Genome Editing in Rat Zygotes.Methods in molecular biology (Clifton, N.J.) · 2023Article
- MosaicVeterinary medicine international · 2023Article
- Pigs with anFrontiers in cell and developmental biology · 2023Article
- Multiple gene editing in porcine embryos using a combination of microinjection, electroporation, and transfection methods.Veterinary world · 2022Article
- Advances in pig models of human diseases.Animal models and experimental medicine · 2022Review
- Zona pellucida treatment before CRISPR/Cas9-mediated genome editing of porcine zygotes.Veterinary medicine and science · 2022Article
- CRISPR/Cas9-editing ofFrontiers in genetics · 2022Article
- Generation of mutant pigs by lipofection-mediated genome editing in embryos.Scientific reports · 2021Article
- Timing and duration of lipofection-mediated CRISPR/Cas9 delivery into porcine zygotes affect gene-editing events.BMC research notes · 2021Article
- Exosome/Liposome-like Nanoparticles: New Carriers for CRISPR Genome Editing in Plants.International journal of molecular sciences · 2021Review
- Current status of the application of gene editing in pigs.The Journal of reproduction and development · 2021Review
- One-Step Generation of Multiple Gene-Edited Pigs by Electroporation of the CRISPR/Cas9 System into Zygotes to Reduce Xenoantigen Biosynthesis.International journal of molecular sciences · 2021Article
- Lipofection-Mediated Introduction of CRISPR/Cas9 System into Porcine Oocytes and Embryos.Animals : an open access journal from MDPI · 2021Article
- Electroporation-Mediated Genome Editing of Livestock Zygotes.Frontiers in genetics · 2021Review
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundXenoantigens are a major source of concern with regard to the success of interspecific xenografts. GGTA1 encodes α1,3-galactosyltransferase, which is essential for the biosynthesis of galactosyl-alpha 1,3-galactose, the major xenoantigen causing hyperacute rejection. GGTA1-modified pigs, therefore, are promising donors for pig-to-human xenotransplantation. In this study, we developed a method for the introduction of the CRISPR/Cas9 system into in vitro-fertilized porcine zygotes via electroporation to generate GGTA1-modified pigs.
resultsWe designed five guide RNAs (gRNAs) targeting distinct sites in GGTA1. After the introduction of the Cas9 protein with each gRNA via electroporation, the gene editing efficiency in blastocysts developed from zygotes was evaluated. The gRNA with the highest gene editing efficiency was used to generate GGTA1-edited pigs. Six piglets were delivered from two recipient gilts after the transfer of electroporated zygotes with the Cas9/gRNA complex. Deep sequencing analysis revealed that five out of six piglets carried a biallelic mutation in the targeted region of GGTA1, with no off-target events. Furthermore, staining with isolectin B4 confirmed deficient GGTA1 function in GGTA1 biallelic mutant piglets.
conclusionsWe established GGTA1-modified pigs with high efficiency by introducing a CRISPR/Cas9 system into zygotes via electroporation. Multiple gene modifications, including knock-ins of human genes, in porcine zygotes via electroporation may further improve the application of the technique in pig-to-human xenotransplantation.
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