ArticleFrontiers in genetics2020
Efficient One-Step Knockout by Electroporation of Ribonucleoproteins Into Zona-Intact Bovine Embryos.
Article in Frontiers in genetics, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 43 citations in OpenAlex.
- Electroporation-mediated site-specific knock-in of a human coding sequence into the bovine ALB locus in embryos.BMC biotechnology · 2026Article
- Selection of two pronuclei zygotes before electroporation does not improve gene editing efficiency or reduce mosaicism in porcine embryos despite optimized timing and Hoechst staining.Veterinary world · 2026Article
- The interleukin-6 signal transducer receptor subunit is required for optimal in vitro bovine embryo development†.Biology of reproduction · 2025Article
- Comparison Between Electroporation at Different Voltage Levels and Microinjection to Generate Porcine Embryos with Multiple Xenoantigen Knock-Outs.International journal of molecular sciences · 2024Article
- Opportunities for CRISPR-Cas9 application in farm animal genetic improvement.Molecular biology reports · 2024Review
- An overview of recent technological developments in bovine genomics.Veterinary and animal science · 2024Review
- Protocol for the electroporation of CRISPR-Cas for DNA and RNA targeting in Bos taurus zygotes.STAR protocols · 2024Article
- Gene editing in livestock: innovations and applications.Animal reproduction · 2024Article
- Livestock embryonic stem cells for reproductive biotechniques and genetic improvement.Animal reproduction · 2024Article
- Ablation ofPNAS nexus · 2023Article
- Optimization of piggyBac Transposon System Electrotransfection in Sheep Fibroblasts.Molecular biotechnology · 2023Article
- Generation of double knockout cattle via CRISPR-Cas9 ribonucleoprotein (RNP) electroporation.Journal of animal science and biotechnology · 2023Article
- Electroporation-Mediated CRISPR/Cas9 Genome Editing in Rat Zygotes.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Optimization of piggyBac transposon-mediated gene transfer method in common marmoset embryos.PloS one · 2023Article
- Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes.Journal of visualized experiments : JoVE · 2022Article
- Towards progressive regulatory approaches for agricultural applications of animal biotechnology.Transgenic research · 2022Review
- CRISPR/Cas9 ribonucleoprotein-mediated genome and epigenome editing in mammalian cells.Advanced drug delivery reviews · 2022Review
- A New Toolbox in Experimental Embryology-Alternative Model Organisms for Studying Preimplantation Development.Journal of developmental biology · 2021Review
- Electroporation-Mediated Genome Editing of Livestock Zygotes.Frontiers in genetics · 2021Review
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Somatic cell nuclear transfer or cytoplasm microinjection have been used to generate genome-edited farm animals; however, these methods have several drawbacks that reduce their efficiency. This study aimed to develop electroporation conditions that allow delivery of CRISPR/Cas9 system to bovine zygotes for efficient gene knock-out. We optimized electroporation conditions to deliver Cas9:sgRNA ribonucleoproteins to bovine zygotes without compromising embryo development. Higher electroporation pulse voltage resulted in increased membrane permeability; however, voltages above 15 V/mm decreased embryo developmental potential. The zona pellucida of bovine embryos was not a barrier to efficient RNP electroporation. Using parameters optimized for maximal membrane permeability while maintaining developmental competence we achieved high rates of gene editing when targeting bovine OCT4, which resulted in absence of OCT4 protein in 100% of the evaluated embryos and the expected arrest of embryonic development at the morula stage. In conclusion, Cas9:sgRNA ribonucleoproteins can be delivered efficiently by electroporation to zona-intact bovine zygotes, resulting in efficient gene knockouts.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.