ArticleScientific reports2019
Strategies to reduce genetic mosaicism following CRISPR-mediated genome edition in bovine embryos.
Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 38 papers, 1 of them a synthesis that pooled it.
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Who cites it
38 citing papers in PubMed, 1 synthesis or guideline pooled it.
- Lineage Differentiation Markers as a Proxy for Embryo Viability in Farm Ungulates.Frontiers in veterinary science · 2021Pooled it
- Resolving Cattle GWAS Loci: Current Progress, Persistent Challenges and Future Directions.Current issues in molecular biology · 2026Review
- Electroporation-mediated site-specific knock-in of a human coding sequence into the bovine ALB locus in embryos.BMC biotechnology · 2026Article
- Targeted Gene Editing in Honey Bees Using Liposome-Based CRISPR-Cas9.Biochemical genetics · 2026Article
- Gene editing of the GJB2 locus in porcine embryos using CRISPR/Cas9 and cytosine base editors: toward a model of congenital deafness.Scientific reports · 2026Article
- Effect of a truncated mutant factor V on hemostatic function and embryonic development in mice.Scientific reports · 2026Article
- Developmental origins of pregnancy loss in cattle.Animal reproduction · 2026Article
- Male gamete copies to characterize genome inheritance and generate progenies.Scientific reports · 2025Article
- CRISPR-Cas9 in the Tailoring of Genetically Engineered Animals.Current issues in molecular biology · 2025Review
- Development of a novel and viable knock-in factor V deficiency murine model: Utility for an ultra-rare disease.PloS one · 2025Article
- SMC2 ablation impairs bovine embryo development shortly after blastocyst hatching.Reproduction (Cambridge, England) · 2024Article
- The role of TEAD4 in trophectoderm commitment and development is not conserved in non-rodent mammals.Development (Cambridge, England) · 2024Article
- PPARG is dispensable for bovine embryo development up to tubular stages†.Biology of reproduction · 2024Article
- Protocol for the electroporation of CRISPR-Cas for DNA and RNA targeting in Bos taurus zygotes.STAR protocols · 2024Article
- Article
- Gene editing in small and large animals for translational medicine: a review.Animal reproduction · 2024Review
- Article
- Ablation ofPNAS nexus · 2023Article
- Highly efficient CRISPR-mediated gene editing in a rotifer.PLoS biology · 2023Article
- Points to consider in the development of national human genome editing policy.Cambridge prisms. Precision medicine · 2023Review
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Genetic mosaicism is the presence of more than two alleles on an individual and it is commonly observed following CRISPR microinjection of zygotes. This phenomenon appears when DNA replication precedes CRISPR-mediated genome edition and it is undesirable because it reduces greatly the odds for direct KO generation by randomly generated indels. In this study, we have developed alternative protocols to reduce mosaicism rates following CRISPR-mediated genome edition in bovine. In a preliminary study we observed by EdU incorporation that DNA replication has already occurred at the conventional microinjection time (20 hpi). Aiming to reduce mosaicism appearance, we have developed three alternative microinjection protocols: early zygote microinjection (10 hpi RNA) or oocyte microinjection before fertilization with either RNA or Ribonucleoprotein delivery (0 hpi RNA or 0 hpi RNP). All three alternative microinjection protocols resulted in similar blastocyst and genome edition rates compared to the conventional 20 hpi group, whereas mosaicism rates were significantly reduced in all early delivery groups (~10-30% of edited embryos being mosaic depending on the loci) compared to conventional 20 hpi microinjection (100% mosaicism rate). These strategies constitute an efficient way to reduce the number of indels, increasing the odds for direct KO generation.
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