ArticleScientific reports2020
Evaluation of mutation rates, mosaicism and off target mutations when injecting Cas9 mRNA or protein for genome editing of bovine embryos.
Article in Scientific reports, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed, 54 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
- 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
- Enhancing the specificity of gene editing outcomes by using Cas9 variants in porcine embryos.Journal of animal science · 2026Article
- Gene, genetics and genetic medicines in gastroenterology: Current status and its future.World journal of gastroenterology · 2026Review
- Analysis of Cytosine Base Editors in Bovine Zygotes: Efficiency and Editing Window Characterization Through Targeting theCurrent issues in molecular biology · 2025Article
- Genome-wide Genetic Mutations Accumulated in Pigs Genome-edited for Xenotransplantation and Their Filial Generation.Genomics, proteomics & bioinformatics · 2025Article
- MSTN gene editing does not alter reproductive capacity but modifies gestational metabolism in Mongolian cattle.BMC veterinary research · 2025Article
- New developments in biotechnology applied to animals: An assessment of the adequacy and sufficiency of current EFSA guidance for animal risk assessment.EFSA journal. European Food Safety Authority · 2025Article
- Immune checkpoint inhibitors in cancer therapy: what lies beyond monoclonal antibodies?Medical oncology (Northwood, London, England) · 2025Review
- The interleukin-6 signal transducer receptor subunit is required for optimal in vitro bovine embryo development†.Biology of reproduction · 2025Article
- Engineered CRISPR-Cas9 for Streptomyces sp. genome editing to improve specialized metabolite production.Nature communications · 2025Article
- Alternatives to antibiotics for sustainable livestock production in the context of the One Health approach: tackling a common foe.Frontiers in veterinary science · 2025Review
- Electroporation Delivery of Cas9 sgRNA Ribonucleoprotein-Mediated Genome Editing in Sheep IVF Zygotes.International journal of molecular sciences · 2024Article
- Unexpected mutations occurred in CRISPR/Cas9 edited Drosophila analyzed by deeply whole genomic sequencing.Heliyon · 2024Article
- Germline ablation achieved via CRISPR/Cas9 targeting ofFrontiers in genome editing · 2023Article
- Research Progress on Nanoparticles-Based CRISPR/Cas9 System for Targeted Therapy of Tumors.Biomolecules · 2022Review
- Article
- LincRNA#1 knockout alone does not affect polled phenotype in cattle heterozygous for the celtic POLLED allele.Scientific reports · 2022Article
- A deletion at the polled PScientific reports · 2022Article
- Cytoplasmic Injection of Zygotes to Genome Edit Naturally Occurring Sequence Variants Into Bovine Embryos.Frontiers in genetics · 2022Article
Corrections and comments
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
Abstract
The CRISPR/Cas9 genome editing tool has the potential to improve the livestock breeding industry by allowing for the introduction of desirable traits. Although an efficient and targeted tool, the CRISPR/Cas9 system can have some drawbacks, including off-target mutations and mosaicism, particularly when used in developing embryos. Here, we introduced genome editing reagents into single-cell bovine embryos to compare the effect of Cas9 mRNA and protein on the mutation efficiency, level of mosaicism, and evaluate potential off-target mutations utilizing next generation sequencing. We designed guide-RNAs targeting three loci (POLLED, H11, and ZFX) in the bovine genome and saw a significantly higher rate of mutation in embryos injected with Cas9 protein (84.2%) vs. Cas9 mRNA (68.5%). In addition, the level of mosaicism was higher in embryos injected with Cas9 mRNA (100%) compared to those injected with Cas9 protein (94.2%), with little to no unintended off-target mutations detected. This study demonstrated that the use of gRNA/Cas9 ribonucleoprotein complex resulted in a high editing efficiency at three different loci in bovine embryos and decreased levels of mosaicism relative to Cas9 mRNA. Additional optimization will be required to further reduce mosaicism to levels that make single-step embryo editing in cattle commercially feasible.
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Registered trials
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