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A mouse knockout library for secreted and transmembrane proteins

Nature Biotechnology volume 28, pages 749–755 (2010)Cite this article

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Abstract

Large collections of knockout organisms facilitate the elucidation of gene functions. Here we used retroviral insertion or homologous recombination to disrupt 472 genes encoding secreted and membrane proteins in mice, providing a resource for studying a large fraction of this important class of drug target. The knockout mice were subjected to a systematic phenotypic screen designed to uncover alterations in embryonic development, metabolism, the immune system, the nervous system and the cardiovascular system. The majority of knockout lines exhibited altered phenotypes in at least one of these therapeutic areas. To our knowledge, a comprehensive phenotypic assessment of a large number of mouse mutants generated by a gene-specific approach has not been described previously.

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Figure 1: Gene selection based on expression.
Figure 2: Summary of phenotypes.
Figure 3: Clec1b deficient mice.

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Acknowledgements

We thank J. Brennan, S. Bunting, L. Corson, P. Fielder, E. Filvaroff, D. French, J. Junutula, F. Peale, H. Phillips, M. Rohrer, H. Stern, J. Zha, R. Watts, B. Wolf and scientists in the Genentech Immunology Department for critical review of the knockout phenotypes and E. Bierwagen and D. Wan for the bioinformatics infrastructures used to track phenotypic calls. We thank J. Mitchell for analysis and plotting of histograms depicting phenotypic ranges. We also thank M. Tessier-Lavigne, F. Bazan, M. Kong-Beltran, J. Theunissen, S. Warming and Z. Zhang for critical reading of the manuscript.

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Authors and Affiliations

  1. Department of Molecular Biology, Genentech, Inc., South San Francisco, California, USA

    Tracy Tang, Deanna Grant, Mark Solloway, Nico Ghilardi, Andrew Peterson & Frederic J de Sauvage

  2. Bioinformatics, Genentech, Inc., South San Francisco, California, USA

    Li Li, Jerry Tang & Yun Li

  3. Immunology, Genentech, Inc., South San Francisco, California, USA

    Wei Yu Lin, Flavius Martin & Paul Godowski

  4. Tumor Biology & Angiogenesis, Genentech, Inc., South San Francisco, California, USA

    Leon Parker & Weilan Ye

  5. Nonclinical Biostatistics, Genentech, Inc., South San Francisco, California, USA

    William Forrest

  6. Lexicon Pharmaceuticals, Inc., The Woodlands, Texas, USA

    Tamas Oravecz, Kenneth A Platt, Dennis S Rice, Gwenn M Hansen, Alejandro Abuin, Derek E Eberhart, Kathleen H Holt & Brian P Zambrowicz

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  1. Tracy Tang
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Contributions

F.J.d.S., T.T., A.P. and B.P.Z. designed the project, analyzed data and wrote the manuscript. F.M. and P.G. designed experiments and analyzed data. J.T. and N.G. contributed to the identification of murine orthologs. K.H.H. and N.G. contributed to the design and verification of targeting strategies. T.O., K.A.P., D.S.R., G.M.H., A.A., D.E.E., K.H.H. and B.P.Z. designed, performed and supervised the knockout generation and phenotype screen. T.T., L.L. and W.F. contributed to the statistical analysis and making the phenotype calls. J.T. and Y.L. implemented the database for public access. L.L. compiled the adult tissue calls. L.P. and W.Y. performed the embryonic in situ hybridization screen. W.Y.L., F.M., D.G. and M.S. performed the follow-up characterization of the Clec1b mutant line.

Corresponding author

Correspondence to Frederic J de Sauvage.

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Competing interests

T.T., L.L., J.T., Y.L., W.Y.L., F.M., D.G., M.S., L.P., W.Y., W.F., N.G., P.G., A.P. and F.J.d.S. are employees of Genentech, a wholly owned subsidiary of Hoffmann-La Roche. T.O., K.A.P., D.S.R., G.M.H., A.A., D.E.E., K.H.H. and B.P.Z. are employees and shareholders of Lexicon Pharmaceuticals.

Supplementary information

Supplementary Text and Figures

Supplementary Tables 1,3,4,5,8 and Supplementary Figs. 1 and 2 (PDF 807 kb)

Supplementary Table 2

Gene symbol (human) (XLS 607 kb)

Supplementary Table 7

Calls from quantitative analysis (XLS 391 kb)

Supplementary Table 6

Calls from non-quantitative assays (XLS 49 kb)

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Tang, T., Li, L., Tang, J. et al. A mouse knockout library for secreted and transmembrane proteins. Nat Biotechnol 28, 749–755 (2010). https://doi.org/10.1038/nbt.1644

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