ArticleBiology open2016
Ultra-superovulation for the CRISPR-Cas9-mediated production of gene-knockout, single-amino-acid-substituted, and floxed mice.
Article in Biology open, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 31 citations in OpenAlex.
- Developmental Potential of In Vitro Fertilization-Derived Mouse Zygotes Following Vitrification: Effects of Superovulation Method.Journal of the American Association for Laboratory Animal Science : JAALAS · 2025Article
- Optimized cesarean techniques, IVF use, and foster strain selection enhance germ-free mouse production efficiency.Scientific reports · 2025Article
- BMP-9 mediates fibroproliferation in fibrodysplasia ossificans progressiva through TGF-β signaling.EMBO molecular medicine · 2025Article
- GDE5/Gpcpd1 activity determines phosphatidylcholine composition in skeletal muscle and regulates contractile force in mice.Communications biology · 2024Article
- Article
- Modification of improved-genome editing via oviductal nucleic acids delivery (i-GONAD)-mediated knock-in in rats.BMC biotechnology · 2021Article
- Response to "Reproducibility of CRISPR-Cas9 methods for generation of conditional mouse alleles: a multi-center evaluation".Genome biology · 2021Article
- Establishment of a conditional Nomo1 mouse model by CRISPR/Cas9 technology.Molecular biology reports · 2020Article
- Review
- Cryopreservation of mouse resources.Laboratory animal research · 2020Review
- Cas9 Ribonucleoprotein Complex Delivery: Methods and Applications for Neuroinflammation.Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology · 2019Review
- Conditional mutagenesis by oligonucleotide-mediated integration of loxP sites in zebrafish.PLoS genetics · 2018Article
- Electroporation-mediated genome editing in vitrified/warmed mouse zygotes created by IVF via ultra-superovulation.Experimental animals · 2018Article
- Efficient mouse genome engineering by CRISPR-EZ technology.Nature protocols · 2018Article
- Untapped Reserves: Controlling Primordial Follicle Growth Activation.Trends in molecular medicine · 2018Review
- Genome-wide Mapping of Off-Target Events in Single-Stranded Oligodeoxynucleotide-Mediated Gene Repair Experiments.Molecular therapy : the journal of the American Society of Gene Therapy · 2018Article
- Generation and Analysis of Striated Muscle Selective LINC Complex Protein Mutant Mice.Methods in molecular biology (Clifton, N.J.) · 2018Article
- Generation of Genetically Modified Mice through the Microinjection of Oocytes.Journal of visualized experiments : JoVE · 2017Article
- Culture time of vitrified/warmed zygotes before microinjection affects the production efficiency of CRISPR-Cas9-mediated knock-in mice.Biology open · 2017Article
- CRISPR/Cas9-mediated gene manipulation to create single-amino-acid-substituted and floxed mice with a cloning-free method.Scientific reports · 2017Article
Corrections and comments
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Authors and funding
8 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Current advances in producing genetically modified mice using genome-editing technologies have indicated the need for improvement of limiting factors including zygote collection for microinjection and their cryopreservation. Recently, we developed a novel superovulation technique using inhibin antiserum and equine chorionic gonadotropin to promote follicle growth. This method enabled the increased production of fertilized oocytes via in vitro fertilization compared with the conventional superovulation method. Here, we verify that the ultra-superovulation technique can be used for the efficient generation of clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9)-mediated knockout mice by microinjection of plasmid vector or ribonucleoprotein into zygotes. We also investigated whether single-amino-acid-substituted mice and conditional knockout mice could be generated. Founder mice bearing base substitutions were generated more efficiently by co-microinjection of Cas9 protein, a guide RNA and single-stranded oligodeoxynucleotide (ssODN) than by plasmid microinjection with ssODN. The conditional allele was successfully introduced by the one-step insertion of an ssODN designed to carry an exon flanked by two loxP sequences and homology arms using a double-cut CRISPR-Cas9 strategy. Our study presents a useful method for the CRISPR-Cas9-based generation of genetically modified mice from the viewpoints of animal welfare and work efficiency.
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Registered trials
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.