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Functional analysis of secreted and transmembrane proteins critical to mouse development

Nature Genetics volume 28, pages 241–249 (2001)Cite this article

Abstract

We describe the successful application of a modified gene-trap approach, the secretory trap, to systematically analyze the functions in vivo of large numbers of genes encoding secreted and membrane proteins. Secretory-trap insertions in embryonic stem cells can be transmitted to the germ line of mice with high efficiency and effectively mutate the target gene. Of 60 insertions analyzed in mice, one-third cause recessive lethal phenotypes affecting various stages of embryonic and postnatal development. Thus, secretory-trap mutagenesis can be used for a genome-wide functional analysis of cell signaling pathways that are critical for normal mammalian development and physiology.

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Figure 1: Predicted structures of 16 new membrane or secreted proteins.
Figure 2: The secretory-trap vector induces null or near null alleles.
Figure 3: Survey of embryonic, neonatal, and postnatal phenotypes.

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Acknowledgements

We thank A. Smith for providing us with the feeder-independent ES cell lines and the Oct4 probe, C. Wright for the Mox1 probe, J. Gladden for his help on the PST9 phenotype analysis, Sheila Avery for help with formatting Fig. 1, and A. Jeske and S. Elson for technical support. Funding for this project was provided by grants to W.C.S. and M.T-L. by a Program in Genomics Applications from the National Heart, Lung, and Blood Institute (NHLBI), grants to W.C.S. from the BBSRC (UK), the Chicago Community Trust, the NICHD, and the March of Dimes, and to M.T.-L. from the NIMH. The ongoing screen is also supported by a Program in Genomics Applications from the NHBLI. K.J.M. was supported by a fellowship from the Jane Coffin Childs Memorial Fund for Medical Research, O.G.K. by a fellowship from the NSF, L.V.G by a fellowship from the Helen Hay Whitney Foundation.. P.A.L. was a Howard Hughes Medical Institute Fellow, and P.T. is a Burroughs Wellcome Fellow of the Life Sciences Research Foundation. X.L. is a postdoctoral associate and M.T.L. an Investigator of the Howard Hughes Medical Institute. W.C.S. is a 1998 Searle Scholar.

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Author notes

  1. Jane Brennan

    Present address: Department of Molecular & Cell Biology, Harvard University, 16 Divinity Avenue, Cambridge, Massachusetts, 02138, USA

  2. Kevin J. Mitchell and Kathy I. Pinson: These authors contributed equally to the work.

Authors and Affiliations

  1. Department of Molecular and Cell Biology, University of California, Berkeley, 94720, California, USA

    Kevin J. Mitchell, Kathy I. Pinson, Olivia G. Kelly, Joel Zupicich, Paul Scherz, Brian J. Avery, Peri Tate, Kariena Dill, Edivinia Pangilinan, Paul Wakenight & William C. Skarnes

  2. Howard Hughes Medical Institute and Department of Anatomy, University of California, San Francisco, 94143, California, USA

    Kevin J. Mitchell, Philip A. Leighton, Lisa V. Goodrich, Xiaowei Lu & Marc Tessier-Lavigne

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Mitchell, K., Pinson, K., Kelly, O. et al. Functional analysis of secreted and transmembrane proteins critical to mouse development. Nat Genet 28, 241–249 (2001). https://doi.org/10.1038/90074

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