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Cited by 1,708CRISPR and Genetic EngineeringAnimal Genetics and ReproductionPluripotent Stem Cells Research

A conditional knockout resource for the genome-wide study of mouse gene function

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Skarnes, William C., Rosen, Barry, West, Anthony P., Koutsourakis, Manousos, Bushell, Wendy, Iyer, Vivek, Mujica, Alejandro O., Thomas, Mark +10 more·2011·Nature
DOI 10.1038/nature10163PMC PMC3572410
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Abstract

Gene targeting in embryonic stem cells has become the principal technology for manipulation of the mouse genome, offering unrivalled accuracy in allele design and access to conditional mutagenesis. To bring these advantages to the wider research community, large-scale mouse knockout programmes are producing a permanent resource of targeted mutations in all protein-coding genes. Here we report the establishment of a high-throughput gene-targeting pipeline for the generation of reporter-tagged, conditional alleles. Computational allele design, 96-well modular vector construction and high-efficiency gene-targeting strategies have been combined to mutate genes on an unprecedented scale. So far, more than 12,000 vectors and 9,000 conditional targeted alleles have been produced in highly germline-competent C57BL/6N embryonic stem cells. High-throughput genome engineering highlighted by this study is broadly applicable to rat and human stem cells and provides a foundation for future genome-wide efforts aimed at deciphering the function of all genes encoded by the mammalian genome.

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  • Cyfip1 knockout in mouse is embryonically lethal.

    “To study the effects of decreased Cyfip1 levels in vivo, we characterized a Cyfip1 KO mouse41 and found that deletion was embryonically lethal.”

    From The autism and schizophrenia associated gene CYFIP1 is critical for the maintenance of dendritic complexity and the stabilization of mature spines (2014) · cites this paper as [41]

    The citation is attached to the Cyfip1 knockout mouse; the citing authors present embryonic lethality as their own finding. The Abstract and Discussion describe a genome-wide knockout resource, but a search for Cyfip1 found no passage identifying a Cyfip1 knockout mouse, and a search for embryonic lethality found no Cyfip1-specific result. Our copy therefore cannot verify the citation's identification of that mouse. (LLM, conf=0.76)

    Experimental — accuracy not yet measured

  • Several mouse coat color mutants are ribosomal proteins, although they are not the ribosomal components enriched here.

    “Pigment cell enrichment of ribosomal components is not surprising, considering several mouse coat color mutants are ribosomal proteins, although they are not the ribosomal components enriched here [50]–[53].”

    From Gene Expression Analysis of Zebrafish Melanocytes, Iridophores, and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin (2013) · cites this paper as [52]

    The Abstract and Discussion describe a resource for generating mouse knockout alleles, not coat-color phenotypes or ribosomal proteins. Searches of our copy for coat-color mutants and ribosomal pigmentation found no passage addressing the attributed relationship; the claim about which components were enriched also cannot be checked against this source. (LLM, conf=0.74)

    Experimental — accuracy not yet measured

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Cites

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Research works printed but not linked: 48

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Authors
Skarnes, W. C., Rosen, B., West, A. P., Koutsourakis, M., Bushell, W., Iyer, V., Mujica, A. O., Thomas, M., Harrow, J., Cox, T., Jackson, D., Severin, J., Biggs, P., Fu, J., Nefedov, M., de Jong, P. J., Stewart, A. F., & Bradley, A.
Affiliations
  • Wellcome Sanger Institute (GB)
  • Biotechnologisches Zentrum (DE)
Corresponding author
Skarnes, William C.
ORCID
Venue
Nature
Publisher
Springer Science and Business Media LLC
Published
2011-06
Volume
474
Issue
7351
Pages
337-342
ISSN
  • 0028-0836 (ISSN-L)
  • 1476-4687
DOI
10.1038/nature10163
PMC
PMC3572410
OpenAlex
W2085544097
Cited by
1,708
References
0
Citations by year
2026: 74 · 2025: 81 · 2024: 101 · 2023: 109 · 2022: 114 · 2021: 142 · 2020: 133 · 2019: 147 · 2018: 116 · 2017: 116 · 2016: 107 · 2015: 107 · 2014: 122 · 2013: 101 · 2012: 103
FWCI
45.88
Type
journal article
Language
en
Open access
Licence
www.springer.com/tdm (tdm)
Locations
  • Nature (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
Funders
  • Wellcome Trust (077188)
  • National Human Genome Research Institute (U01HG004080)
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
conditional knockout mice, conditional alleles, gene targeting, embryonic stem cells
MeSH terms
Alleles, Animals, Genes, Genes, Lethal, Genetic Vectors, Genotype, Humans, Mice, Inbred C57BL, Phenotype, Polymerase Chain Reaction, Mutagenesis, Insertional, Genome, Gene Deletion, Mice, Knockout, Computational Biology, Genomics, Mice, Rats, Embryonic Stem Cells, Gene Knockout Techniques, Genetic Association Studies

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