ArticleScientific reports2020
Electroporation and genetic supply of Cas9 increase the generation efficiency of CRISPR/Cas9 knock-in alleles in C57BL/6J mouse zygotes.
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 24 papers.
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Who cites it
24 citing papers in PubMed, 42 citations in OpenAlex.
- Cas9-expressing cattle using the PiggyBac transposon all-in-one system.BMC genomics · 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
- Unlocking Genome Editing: Advances and Obstacles in CRISPR/Cas Delivery Technologies.Journal of functional biomaterials · 2024Review
- CRISPR/Cas9 Technology for Enhancing Desirable Traits of Fish Species in Aquaculture.International journal of molecular sciences · 2024Review
- An oocyte-specific Cas9-expressing mouse for germline CRISPR/Cas9-mediated genome editing.Genesis (New York, N.Y. : 2000) · 2024Article
- Increasing Knockin Efficiency in Mouse Zygotes by Transient Hypothermia.The CRISPR journal · 2024Article
- CRISPR/Cas9-Mediated Customizing Strategies for Adoptive T-Cell Therapy.Pharmaceutics · 2024Review
- Donor template delivery by recombinant adeno-associated virus for the production of knock-in mice.BMC biology · 2024Article
- Optimising Electroporation Condition for CRISPR/Cas-Mediated Knockout in Zona-Intact Buffalo Zygotes.Animals : an open access journal from MDPI · 2023Article
- A Review of CRISPR Tools for Treating Usher Syndrome: Applicability, Safety, Efficiency, and In Vivo Delivery.International journal of molecular sciences · 2023Review
- Research and Therapeutic Approaches in Stem Cell Genome Editing by CRISPR Toolkit.Molecules (Basel, Switzerland) · 2023Review
- CRISMERE Chromosome Engineering in Mouse and Rat.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Efficient DNA knock-in using AAV-mediated delivery with 2-cell embryo CRISPR-Cas9 electroporation.Frontiers in genome editing · 2023Article
- Protein structure determination in human cells by in-cell NMR and a reporter system to optimize protein delivery or transexpression.Communications biology · 2022Article
- Highly reliable creation of floxed alleles by electroporating single-cell embryos.BMC biology · 2022Article
- CRISPR/Cas9 ribonucleoprotein-mediated genome and epigenome editing in mammalian cells.Advanced drug delivery reviews · 2022Review
- High-content CRISPR screening in tumor immunology.Frontiers in immunology · 2022Review
- Delivering the CRISPR/Cas9 system for engineering gene therapies: Recent cargo and delivery approaches for clinical translation.Frontiers in bioengineering and biotechnology · 2022Review
- Multidrug-Resistant Microbial Therapy Using Antimicrobial Peptides and the CRISPR/Cas9 System.Veterinary medicine (Auckland, N.Z.) · 2022Review
- Delivery of CRISPR-Cas9 system for screening and editing RNA binding proteins in cancer.Advanced drug delivery reviews · 2022Review
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
Abstract
CRISPR/Cas9 machinery delivered as ribonucleoprotein (RNP) to the zygote has become a standard tool for the development of genetically modified mouse models. In recent years, a number of reports have demonstrated the effective delivery of CRISPR/Cas9 machinery via zygote electroporation as an alternative to the conventional delivery method of microinjection. In this study, we have performed side-by-side comparisons of the two RNP delivery methods across multiple gene loci and conclude that electroporation compares very favourably with conventional pronuclear microinjection, and report an improvement in mutagenesis efficiency when delivering CRISPR via electroporation for the generation of simple knock-in alleles using single-stranded oligodeoxynucleotide (ssODN) repair templates. In addition, we show that the efficiency of knock-in mutagenesis can be further increased by electroporation of embryos derived from Cas9-expressing donor females. The maternal supply of Cas9 to the zygote avoids the necessity to deliver the relatively large Cas9 protein, and high efficiency generation of both indel and knock-in allele can be achieved by electroporation of small single-guide RNAs and ssODN repair templates alone. Furthermore, electroporation, compared to microinjection, results in a higher rate of embryo survival and development. The method thus has the potential to reduce the number of animals used in the production of genetically modified mouse models.
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