The DOI matches its Crossref record; 10 of the 10 references it cites were found in a registry or an index.
Highly Efficient Mouse Genome Editing by CRISPR Ribonucleoprotein Electroporation of Zygotes
Open accessAbstract
The CRISPR/Cas9 system has been employed to efficiently edit the genomes of diverse model organisms. CRISPR-mediated mouse genome editing is typically accomplished by microinjection of Cas9 DNA/RNA and single guide RNA (sgRNA) into zygotes to generate modified animals in one step. However, microinjection is a technically demanding, labor-intensive, and costly procedure with poor embryo viability. Here, we describe a simple and economic electroporation-based strategy to deliver Cas9/sgRNA ribonucleoproteins into mouse zygotes with 100% efficiency for in vivo genome editing. Our methodology, designated as CRISPR RNP Electroporation of Zygotes (CRISPR-EZ), enables highly efficient and high-throughput genome editing in vivo, with a significant improvement in embryo viability compared with microinjection. Using CRISPR-EZ, we generated a variety of editing schemes in mouse embryos, including indel (insertion/deletion) mutations, point mutations, large deletions, and small insertions. In a proof-of-principle experiment, we used CRISPR-EZ to target the tyrosinase (Tyr) gene, achieving 88% bi-allelic editing and 42% homology-directed repair-mediated precise sequence modification in live mice. Taken together, CRISPR-EZ is simple, economic, high throughput, and highly efficient with the potential to replace microinjection for in vivo genome editing in mice and possibly in other mammals.
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Cites
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Cited by
Delivering CRISPR: a review of the challenges and approaches
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Genetic Dissection of Neural Circuits: A Decade of Progress
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Non-viral delivery systems for CRISPR/Cas9-based genome editing: challenges and opportunities
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Metadata
- Authors
- Chen, S., Lee, B., Lee, A. Y.-F., Modzelewski, A. J., & He, L.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2016-07
- Volume
- 291
- Issue
- 28
- Pages
- 14457-14467
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2345889524
- Cited by
- 333
- References
- 10
- FWCI
- 14.30
- Type
- journal article
- Language
- en
- Open access
- hybrid
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- Chen, S., Lee, B., Lee, A. Y.-F., Modzelewski, A. J., & He, L. (2016). Highly Efficient Mouse Genome Editing by CRISPR Ribonucleoprotein Electroporation of Zygotes. Journal of Biological Chemistry. https://doi.org/10.1074/jbc.m116.733154
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