ArticleTransgenic research2019
Introducing gene deletions by mouse zygote electroporation of Cas12a/Cpf1.
Article in Transgenic research, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 28 citations in OpenAlex.
- Ubinuclein 2 is essential for mouse development and functions in X chromosome inactivation.PLoS genetics · 2025Article
- Efficient genome editing of two-cell mouse embryos via modified CRISPR/Cas electroporation.Scientific reports · 2024Article
- Genome editing using type I-E CRISPR-Cas3 in mice and rat zygotes.Cell reports methods · 2024Article
- Epigenetic regulation limits competence of pluripotent stem cell-derived oocytes.The EMBO journal · 2023Article
- Modern Approaches to Mouse Genome Editing Using the CRISPR-Cas Toolbox and Their Applications in Functional Genomics and Translational Research.Advances in experimental medicine and biology · 2023Review
- Amount of Cas9 protein introduced into mouse embryos via electroporation affects the genome-editing rate.The Journal of reproduction and development · 2022Article
- Optimization of Nuclear Localization Signal Composition Improves CRISPR-Cas12a Editing Rates in Human Primary Cells.GEN biotechnology · 2022Article
- RNA-GuidedInternational journal of molecular sciences · 2022Article
- Gene Editing for Inherited Red Blood Cell Diseases.Frontiers in physiology · 2022Review
- Electroporation-Mediated Genome Editing of Livestock Zygotes.Frontiers in genetics · 2021Review
- The genome editing revolution: review.Journal, genetic engineering & biotechnology · 2020Review
- Production of genome-edited mice by visualization of nucleases introduced into the embryos using electroporation.The Journal of reproduction and development · 2020Article
- A most formidable arsenal: genetic technologies for building a better mouse.Genes & development · 2020Review
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
CRISPR-associated (Cas) nucleases are established tools for engineering of animal genomes. These programmable RNA-guided nucleases have been introduced into zygotes using expression vectors, mRNA, or directly as ribonucleoprotein (RNP) complexes by different delivery methods. Whereas microinjection techniques are well established, more recently developed electroporation methods simplify RNP delivery but can provide less consistent efficiency. Previously, we have designed Cas12a-crRNA pairs to introduce large genomic deletions in the Ubn1, Ubn2, and Rbm12 genes in mouse embryonic stem cells (ESC). Here, we have optimized the conditions for electroporation of the same Cas12a RNP pairs into mouse zygotes. Using our protocol, large genomic deletions can be generated efficiently by electroporation of zygotes with or without an intact zona pellucida. Electroporation of as few as ten zygotes is sufficient to obtain a gene deletion in mice suggesting potential applicability of this method for species with limited availability of zygotes.
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