ArticleNucleic acids research2013
Efficient generation of large-scale genome-modified mice using gRNA and CAS9 endonuclease.
Article in Nucleic acids research, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 124 papers, 1 of them a synthesis that pooled it.
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Who cites it
124 citing papers in PubMed, 1 synthesis or guideline pooled it.
- 2015 Guidelines for Establishing Genetically Modified Rat Models for Cardiovascular Research.Journal of cardiovascular translational research · 2015Guideline
- A streamlined approach for gene editing in non-obese diabetes (NOD) mice via CRISPR/Cas9.Laboratory animal research · 2026Article
- Polycomb chromatin topology enables long-range enhancer recruitment during craniofacial development.Nature communications · 2026Article
- Optimized dual-AAV base editor delivery system with enhanced editing efficiency and virion production titer.Synthetic and systems biotechnology · 2025Article
- Newborn Intravenous Injection of Liposomal CRISPR/Cas9 Complex Has No Incidence of Off-Targets or Tumors in Mice.Pharmaceutics · 2025Article
- Deletion of IRS-1 leads to growth failure and insulin resistance with downregulation of liver and muscle insulin signaling in rats.Scientific reports · 2025Article
- Artificial Insemination as a Possible Convenient Tool to Acquire Genome-Edited Mice via In Vivo Fertilization with Engineered Sperm.Biotech (Basel (Switzerland)) · 2024Review
- Chromatin structure in totipotent mouse early preimplantation embryos.The Journal of reproduction and development · 2024Review
- Mutation of the Thap4 gene causes dwarfism and testicular anomalies in rats and mice.Mammalian genome : official journal of the International Mammalian Genome Society · 2024Article
- Establishment of African pygmy mouse induced pluripotent stem cells using defined doxycycline inducible transcription factors.Scientific reports · 2024Article
- Article
- CRISPR/Cas9: an advanced platform for root and tuber crops improvement.Frontiers in genome editing · 2023Review
- Two acquired mouse Y chromosome-linked genes, Prssly and Teyorf1, are dispensable for male fertility‡.Biology of reproduction · 2022Article
- Mice blocking Ser347 phosphorylation of pregnane x receptor develop hepatic fasting-induced steatosis and hypertriglyceridemia.Biochemical and biophysical research communications · 2022Article
- Multiplexed high-throughput localized electroporation workflow with deep learning-based analysis for cell engineering.Science advances · 2022Article
- Speciation and adaptation research meets genome editing.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2022Review
- Production of Germ Cell-Less Rainbow Trout by dead end Gene Knockout and their Use as Recipients for Germ Cell Transplantation.Marine biotechnology (New York, N.Y.) · 2022Article
- Somatostatin-evoked Aβ catabolism in the brain: Mechanistic involvement of α-endosulfine-KMolecular psychiatry · 2022Article
- Recent Advances in the Production of Genome-Edited Rats.International journal of molecular sciences · 2022Review
- Removal of a 10-kbHorticulture research · 2022Article
64 more citing papers are in PubMed but not listed here.
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The generation of genome-modified animals is a powerful approach to analyze gene functions. The CAS9/guide RNA (gRNA) system is expected to become widely used for the efficient generation of genome-modified animals, but detailed studies on optimum conditions and availability are limited. In the present study, we attempted to generate large-scale genome-modified mice with an optimized CAS9/gRNA system, and confirmed the transmission of these mutations to the next generations. A comparison of different types of gRNA indicated that the target loci of almost all pups were modified successfully by the use of long-type gRNAs with CAS9. We showed that this system has much higher mutation efficiency and much lower off-target effect compared to zinc-finger nuclease. We propose that most of these off-target effects can be avoided by the careful control of CAS9 mRNA concentration and that the genome-modification efficiency depends rather on the gRNA concentration. Under optimized conditions, large-scale (~10 kb) genome-modified mice can be efficiently generated by modifying two loci on a single chromosome using two gRNAs at once in mouse zygotes. In addition, the normal transmission of these CAS9/gRNA-induced mutations to the next generation was confirmed. These results indicate that CAS9/gRNA system can become a highly effective tool for the generation of genome-modified animals.
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