ArticlePloS one2015
Detailed phenotypic and molecular analyses of genetically modified mice generated by CRISPR-Cas9-mediated editing.
Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
What it found
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Who cites it
35 citing papers in PubMed, 56 citations in OpenAlex.
- Article
- CRISPR/Cas9-Engineering for Increased Amylolytic Potential of Microbes for Sustainable Wastewater Treatment: A Review.Current microbiology · 2024Review
- Expression of a single inhibitory member of the Ly49 receptor family is sufficient to license NK cells for effector functions.bioRxiv : the preprint server for biology · 2024Article
- Inherited C-terminal TREX1 variants disrupt homology-directed repair to cause senescence and DNA damage phenotypes in Drosophila, mice, and humans.Nature communications · 2024Article
- Various repair events following CRISPR/Cas9-based mutational correction of an infertility-related mutation in mouse embryos.Journal of assisted reproduction and genetics · 2024Article
- Article
- From amputations to antibiotics: A future beyond "hacksaw" gene editing.Molecular therapy : the journal of the American Society of Gene Therapy · 2022Article
- CRISPRthripsis: The Risk of CRISPR/Cas9-induced Chromothripsis in Gene Therapy.Stem cells translational medicine · 2022Article
- Development and characterization of type I interferon receptor knockout sheep: A model for viral immunology and reproductive signaling.Frontiers in genetics · 2022Article
- Novel Mouse Model Reveals That Serine Phosphorylation of L-Plastin Is Essential for Effective Splenic Clearance of Pneumococcus.Journal of immunology (Baltimore, Md. : 1950) · 2021Article
- DGK and DZHK position paper on genome editing: basic science applications and future perspective.Basic research in cardiology · 2021Review
- Forward and Reverse Genetics of B Cell Malignancies: From Insertional Mutagenesis to CRISPR-Cas.Frontiers in immunology · 2021Review
- Control of Viral Infection by Natural Killer Cell Inhibitory Receptors.Cell reports · 2020Article
- The technical risks of human gene editing.Human reproduction (Oxford, England) · 2019Article
- Microhomologies are prevalent at Cas9-induced larger deletions.Nucleic acids research · 2019Article
- Off- and on-target effects of genome editing in mouse embryos.The Journal of reproduction and development · 2019Article
- Gene-edited CRISPy Critters for alcohol research.Alcohol (Fayetteville, N.Y.) · 2019Review
- Genome Editing in Mice Using CRISPR/Cas9 Technology.Current protocols in cell biology · 2018Article
- Re-evaluation of mouse tissue factor pathway inhibitor and comparison of mouse and human tissue factor pathway inhibitor physiology.Journal of thrombosis and haemostasis : JTH · 2018Article
- Repair of double-strand breaks induced by CRISPR-Cas9 leads to large deletions and complex rearrangements.Nature biotechnology · 2018Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
The bacterial CRISPR-Cas9 system has been adapted for use as a genome editing tool. While several recent reports have indicated that successful genome editing of mice can be achieved, detailed phenotypic and molecular analyses of the mutant animals are limited. Following pronuclear micro-injection of fertilized eggs with either wild-type Cas9 or the nickase mutant (D10A) and single or paired guide RNA (sgRNA) for targeting of the tyrosinase (Tyr) gene, we assessed genome editing in mice using rapid phenotypic readouts (eye and coat color). Mutant mice with insertions or deletions (indels) in Tyr were efficiently generated without detectable off-target cleavage events. Gene correction of a single nucleotide by homologous recombination (HR) could only occur when the sgRNA recognition sites in the donor DNA were modified. Gene repair did not occur if the donor DNA was not modified because Cas9 catalytic activity was completely inhibited. Our results indicate that allelic mosaicism can occur following -Cas9-mediated editing in mice and appears to correlate with sgRNA cleavage efficiency at the single-cell stage. We also show that larger than expected deletions may be overlooked based on the screening strategy employed. An unbiased analysis of all the deleted nucleotides in our experiments revealed that the highest frequencies of nucleotide deletions were clustered around the predicted Cas9 cleavage sites, with slightly broader distributions than expected. Finally, additional analysis of founder mice and their offspring indicate that their general health, fertility, and the transmission of genetic changes were not compromised. These results provide the foundation to interpret and predict the diverse outcomes following CRISPR-Cas9-mediated genome editing experiments in mice.
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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.