ArticleTransgenic research2017
Efficient gene targeting in mouse zygotes mediated by CRISPR/Cas9-protein.
Article in Transgenic research, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Gene Therapy for Inherited Liver Disease: To Add or to Edit.International journal of molecular sciences · 2024Review
- Embryo and fetal gene editing: Technical challenges and progress toward clinical applications.Molecular therapy. Methods & clinical development · 2024Review
- Donor template delivery by recombinant adeno-associated virus for the production of knock-in mice.BMC biology · 2024Article
- Genetically modified mice as a tool for the study of human diseases.Molecular biology reports · 2024Review
- The International Mouse Phenotyping Consortium: comprehensive knockout phenotyping underpinning the study of human disease.Nucleic acids research · 2023Article
- Advances in Genome Editing and Application to the Generation of Genetically Modified Rat Models.Frontiers in genetics · 2021Review
- Review
- Introducing gene deletions by mouse zygote electroporation of Cas12a/Cpf1.Transgenic research · 2019Article
- Genetic cartography of longevity in humans and mice: Current landscape and horizons.Biochimica et biophysica acta. Molecular basis of disease · 2018Review
- Comparative analysis of single-stranded DNA donors to generate conditional null mouse alleles.BMC biology · 2018Article
- Efficient mouse genome engineering by CRISPR-EZ technology.Nature protocols · 2018Article
- Long Evans rat spermatogonial lines are effective germline vectors for transgenic rat production.Transgenic research · 2017Article
- Quantitative assessment of timing, efficiency, specificity and genetic mosaicism of CRISPR/Cas9-mediated gene editing of hemoglobin beta gene in rhesus monkey embryos.Human molecular genetics · 2017Article
- Easi-CRISPR: a robust method for one-step generation of mice carrying conditional and insertion alleles using long ssDNA donors and CRISPR ribonucleoproteins.Genome biology · 2017Article
- HMEJ-based safe-harbor genome editing enables efficient generation of cattle with increased resistance to tuberculosis.The Journal of biological chemistryArticle
Corrections and comments
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Authors and funding
7 authors.
Funding
Abstract
The CRISPR/Cas9 system has rapidly advanced targeted genome editing technologies. However, its efficiency in targeting with constructs in mouse zygotes via homology directed repair (HDR) remains low. Here, we systematically explored optimal parameters for targeting constructs in mouse zygotes via HDR using mouse embryonic stem cells as a model system. We characterized several parameters, including single guide RNA cleavage activity and the length and symmetry of homology arms in the construct, and we compared the targeting efficiency between Cas9, Cas9nickase, and dCas9-FokI. We then applied the optimized conditions to zygotes, delivering Cas9 as either mRNA or protein. We found that Cas9 nucleo-protein complex promotes highly efficient, multiplexed targeting of circular constructs containing reporter genes and floxed exons. This approach allows for a one-step zygote injection procedure targeting multiple genes to generate conditional alleles via homologous recombination, and simultaneous knockout of corresponding genes in non-targeted alleles via non-homologous end joining.
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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.