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

By W. Skarnes, B. Rosen, A. West, M. Koutsourakis, Wendy Bushell, V. Iyer, A. Mujica, Mark Thomas, J. Harrow, Tony Cox and 8 more. Nature, 2011

1,720 citations recorded by the source corpus.

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.

Subfields: Genetics, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/f4af7779667d6fb0aa3c014054fc6510ad197344

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Citations in the corpus

RCSB Protein Data Bank: powerful new tools for exploring 3D structures of biological macromolecules for basic and applied research and education in fundamental biology, biomedicine, biotechnology, bioengineering and energy sciences (2020, background): “org (51)] and Pharos––Illuminating the Druggable Genome [https://pharos.”
Synergism of TNF-α and IFN-γ Triggers Inflammatory Cell Death, Tissue Damage, and Mortality in SARS-CoV-2 Infection and Cytokine Shock Syndromes (2020, neutral): “Gsdmd–/–Gsdme–/–(Gsdmd/ e–/–)Mlkl–/– mice were generated at our facility by crossingGsdme–/–,Gsdmd–/–, andMlkl–/– mice.”
The PR/SET Domain Zinc Finger Protein Prdm4 Regulates Gene Expression in Embryonic Stem Cells but Plays a Nonessential Role in the Developing Mouse Embryo (2013, methodology): “To confirm that Prdm4 is dispensable for mouse development, we also generated a second targeted allele using the EUCOMM/ KOMP resource (68).”
High-throughput discovery of novel developmental phenotypes (2016, positive): “Taking advantage of the LacZ cassette present in most IMPC alleles 10,11 , gene expression was evaluated in heterozygous embryos at E12.”
Cleavage of DFNA5 by caspase-3 during apoptosis mediates progression to secondary necrotic/pyroptotic cell death (2017, positive): “These mice were generated from the KOMP/EUCOMM targeted ES cell resource using standard techniques as described previously 60,61 and established on a C57BL/6 genetic background.”
Computational Drug Repositioning: From Data to Therapeutics (2013, background): “41 Surprisingly, little or no use of knockout data has yet been made to directly and systematically reposition drugs.”
DUSP11-mediated control of 5′-triphosphate RNA regulates RIG-I sensitivity (2020, background): “The knockout first (Skarnes et al. 2011) Dusp11tm1a(EUCOMM)Wtsi mutation encodes a splicing acceptor (SA) following Dusp11 exon 1 that disrupts targeted gene expression (Supplemental Fig.”
Secreted Metalloproteinase ADAMTS-3 Inactivates Reelin (2017, methodology): “Knockout-First ADAMTS3 heterozygous mice Project ID CSD50174: Adamts3 tm1a(KOMP)Wtsi; information is available at https:// www.komp.org/ were obtained from the Knockout Mouse Project (KOMP) Repository/Welcome Trust Sanger Institute (Skarnes et al., 2011; Bradley et al., 2012).”
The Nerve Growth Factor Signaling and Its Potential as Therapeutic Target for Glaucoma (2014, methodology): “So far, a conditional knockout mouse line of OPTN has been generated and is available at the Wellcome Trust Sanger Institute [87]; Myoc-null mice have also been produced by Kim and coworkers [88]; these genetically altered mice are useful tools for understanding the physiological function of glaucom”