ArticleScientific reports2015
Generation of mutant mice via the CRISPR/Cas9 system using FokI-dCas9.
Article in Scientific reports, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed.
- Identification of maternal allele sequences of IG-DMR that are essential for neonatal viability.PloS one · 2025Article
- Microinjection of the CRISPR/Cas9 editing system through the germ pore of a wheat microspore induces mutations in the target Ms2 gene.Molecular biology reports · 2024Article
- CRISPR/Cpf1-FOKI-induced gene editing inSynthetic and systems biotechnology · 2024Article
- CRISPR-Based Genome Editing Tools: An Accelerator in Crop Breeding for a Changing Future.International journal of molecular sciences · 2023Review
- TEAD1 trapping by the Q353R-Lamin A/C causes dilated cardiomyopathy.Science advances · 2023Article
- Recent Advances in Improving Gene-Editing Specificity through CRISPR-Cas9 Nuclease Engineering.Cells · 2022Review
- Review
- Application of Various Delivery Methods for CRISPR/dCas9.Molecular biotechnology · 2020Review
- Article
- Efficient production and transmission of CRISPR/Cas9-mediated mutant alleles at the IG-DMR via generation of mosaic mice using a modified 2CC method.Scientific reports · 2019Article
- An update on the tools for creating transgenic animal models of human diseases - focus on atherosclerosis.Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologica · 2019Review
- High-fidelity endonuclease variant HypaCas9 facilitates accurate allele-specific gene modification in mouse zygotes.Communications biology · 2019Article
- Mapping of a responsible region for sex reversal upstream of Sox9 by production of mice with serial deletion in a genomic locus.Scientific reports · 2018Article
- Collagen-Induced Arthritis Analysis inBiomolecules & therapeutics · 2018Article
- Targeted insertion of an anti-CD2 monoclonal antibody transgene into the GGTA1 locus in pigs using FokI-dCas9.Scientific reports · 2017Article
- CRISPR-Cas orthologues and variants: optimizing the repertoire, specificity and delivery of genome engineering tools.Mammalian genome : official journal of the International Mammalian Genome Society · 2017Review
- CRISPR/Cas9-mediated simultaneous knockout of Dmrt1 and Dmrt3 does not recapitulate the 46,XY gonadal dysgenesis observed in 9p24.3 deletion patients.Biochemistry and biophysics reports · 2017Article
- CAR1 deletion by CRISPR/Cas9 reduces formation of ethyl carbamate from ethanol fermentation by Saccharomyces cerevisiae.Journal of industrial microbiology & biotechnology · 2016Article
- CRISPR RNA-guided FokI nucleases repair a PAH variant in a phenylketonuria model.Scientific reports · 2016Article
- Microinjection-based generation of mutant mice with a double mutation and a 0.5 Mb deletion in their genome by the CRISPR/Cas9 system.The Journal of reproduction and development · 2016Article
Corrections and comments
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Authors and funding
9 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Genome editing, which introduces mutations in genes of interest using artificial DNA nucleases such as the ZFN, TALEN, and CRISPR/Cas9 systems in living cells, is a useful tool for generating mutant animals. Although CRISPR/Cas9 provides advantages over the two other systems, such as an easier vector construction and high efficiency of genome editing, it raises concerns of off-target effects when single guide RNA (gRNA) is used. Recently, FokI-dCas9 (fCas9), a fusion protein comprised of the inactivated mutant form of Cas9 and the DNA nuclease domain of FokI, has been developed. It enables genome editing with reduced risks of off-target effects in mammalian cultured cell lines, as fCas9 requires gRNAs to bind opposite strands with an appropriate distance between them. Here, we demonstrated that fCas9 efficiently generates living mutant mice through microinjection of its mRNA and gRNAs into zygotes. A comparison of the relative efficiencies of genome editing using fCas9 and other modified Cas9s showed that these mutagenesis efficiencies are similar when the targets of two gRNAs are separated by an appropriate distance, suggesting that in addition to the ease of vector construction, fCas9 exhibit high efficiency in producing mutant mice and in reducing risks of off-target effects.
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Registered trials
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