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Somatic mosaicism and allele complexity induced by CRISPR/Cas9 RNA injections in mouse zygotes
Open accessAbstract
Tyrosinase is the rate-limiting enzyme for the production of melanin pigmentation. In the mouse and other animals, homozygous null mutations in the Tyrosinase gene (Tyr) result in the absence of pigmentation, i.e. albinism. Here we used the CRISPR/Cas9 system to generate mono- and bi-allelic null mutations in the Tyr locus by zygote injection of two single-guide and Cas9 RNAs. Injection into C57BL/6N wild-type embryos resulted in one completely albino founder carrying two different Tyr mutations. In addition, three pigmentation mosaics and fully pigmented littermates were obtained that transmitted new mutant Tyr alleles to progeny in test crosses with albinos. Injection into Tyr heterozygous (B6CBAF1/J×FVB/NJ) zygotes resulted in the generation of numerous albinos and also mice with a graded range of albino mosaicism. Deep sequencing revealed that the majority of the albinos and the mosaics had more than two new mutant alleles. These visual phenotypes and molecular genotypes highlight the somatic mosaicism and allele complexity in founders that occurs for targeted genes during CRISPR/Cas9-mediated mutagenesis by zygote injection in mice.
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“We expect incomplete penetrance of the perturbation in this dataset because of the mosaic nature of Cas9 mutagenesis29.”
From Quantifying the effect of experimental perturbations at single-cell resolution (2021) · cites this paper as [29]
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Metadata
- Authors
- Yen, S.-T., Zhang, M., Deng, J. M., Usman, S. J., Smith, C. N., Parker-Thornburg, J., Swinton, P. G., Martin, J. F., & Behringer, R. R.
- Affiliations
- The University of Texas MD Anderson Cancer Center (US)
- Baylor College of Medicine (US)
- Corresponding author
- Behringer, Richard R.
- Venue
- Developmental Biology
- Publisher
- Elsevier BV
- Published
- 2014-09
- Volume
- 393
- Issue
- 1
- Pages
- 3-9
- ISSN
- 0012-1606 (ISSN-L)
- PMC
- PMC4166609
- OpenAlex
- W2089929813
- Cited by
- 300
- References
- 40
- Citations by year
- 2026: 15 · 2025: 4 · 2024: 19 · 2023: 20 · 2022: 21 · 2021: 22 · 2020: 30 · 2019: 40 · 2018: 31 · 2017: 36 · 2016: 32 · 2015: 21 · 2014: 9
- FWCI
- 13.40
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Developmental Biology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health (DE023177, CA016672)
- Smith Foundation to J.F.M. and National Institutes of Health (HD030284)
- University of Texas MD Anderson Genetics
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Keywords
- somatic mosaicism, CRISPR/Cas9, zygote injection, tyrosinase, TYR gene, albinism, melanin pigmentation, targeted mutagenesis, mouse embryo
- MeSH terms
- Gene Editing, CRISPR-Associated Protein 9, Albinism, Alleles, Animals, Bacterial Proteins, Base Sequence, Endonucleases, Female, Male, Melanins, Mice, Inbred C57BL, Mosaicism, Mutation, Pigmentation, RNA, Messenger, Monophenol Monooxygenase, Zygote, Mutagenesis, Sequence Analysis, DNA, Mice, RNA, Small Untranslated, High-Throughput Nucleotide Sequencing, CRISPR-Cas Systems, CRISPR-Associated Proteins
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