Cited by 300CRISPR and Genetic EngineeringAnimal Genetics and ReproductionPluripotent Stem Cells Research

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Somatic mosaicism and allele complexity induced by CRISPR/Cas9 RNA injections in mouse zygotes

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Yen, Shuo-Ting, Zhang, Min, Deng, Jian Min, Usman, Shireen J., Smith, Chad N., Parker-Thornburg, Jan, Swinton, Paul G., Martin, James F., Behringer, Richard R.·2014·Developmental Biology

DOI 10.1016/j.ydbio.2014.06.017PMC PMC4166609

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Abstract

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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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.
ORCID: Yen, Shuo-Ting
Zhang, Min
Deng, Jian Min
Martin, James F.
Behringer, Richard R.
Venue: Developmental Biology
Publisher: Elsevier BV
Published: 2014-09
Volume: 393
Issue: 1
Pages: 3-9
ISSN: 0012-1606 (ISSN-L)
DOI: 10.1016/j.ydbio.2014.06.017
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: www.elsevier.com/tdm/userlicense/1.0/ (tdm)
www.elsevier.com/open-access/userlicense/1.0/ (vor, 365 d)
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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Somatic mosaicism and allele complexity induced by CRISPR/Cas9 RNA injections in mouse zygotes

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