Abstract
A strategy based on the gene trap was developed to prescreen mouse embryonic stem cells for insertional mutations in genes encoding secreted and membrane-spanning proteins. The "secretory trap" relies on capturing the N-terminal signal sequence of an endogenous gene to generate an active beta-galactosidase fusion protein. Insertions were found in a cadherin gene, an unc6-related laminin (netrin) gene, the sek receptor tyrosine kinase gene, and genes encoding two receptor-linked protein-tyrosine phosphatases, LAR and PTP kappa. Analysis of homozygous mice carrying insertions in LAR and PTP kappa showed that both genes were effectively disrupted, but neither was essential for normal embryonic development.
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- Beddington R. S., Morgernstern J., Land H., Hogan A. An in situ transgenic enzyme marker for the midgestation mouse embryo and the visualization of inner cell mass clones during early organogenesis. Development. 1989 May;106(1):37–46. doi: 10.1242/dev.106.1.37. [DOI] [PubMed] [Google Scholar]
- Brenner D. G., Lin-Chao S., Cohen S. N. Analysis of mammalian cell genetic regulation in situ by using retrovirus-derived "portable exons" carrying the Escherichia coli lacZ gene. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5517–5521. doi: 10.1073/pnas.86.14.5517. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Burns N., Grimwade B., Ross-Macdonald P. B., Choi E. Y., Finberg K., Roeder G. S., Snyder M. Large-scale analysis of gene expression, protein localization, and gene disruption in Saccharomyces cerevisiae. Genes Dev. 1994 May 1;8(9):1087–1105. doi: 10.1101/gad.8.9.1087. [DOI] [PubMed] [Google Scholar]
- Clark S. J., Jefferies W. A., Barclay A. N., Gagnon J., Williams A. F. Peptide and nucleotide sequences of rat CD4 (W3/25) antigen: evidence for derivation from a structure with four immunoglobulin-related domains. Proc Natl Acad Sci U S A. 1987 Mar;84(6):1649–1653. doi: 10.1073/pnas.84.6.1649. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Friedrich G., Soriano P. Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice. Genes Dev. 1991 Sep;5(9):1513–1523. doi: 10.1101/gad.5.9.1513. [DOI] [PubMed] [Google Scholar]
- Frohman M. A., Dush M. K., Martin G. R. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8998–9002. doi: 10.1073/pnas.85.23.8998. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gossler A., Joyner A. L., Rossant J., Skarnes W. C. Mouse embryonic stem cells and reporter constructs to detect developmentally regulated genes. Science. 1989 Apr 28;244(4903):463–465. doi: 10.1126/science.2497519. [DOI] [PubMed] [Google Scholar]
- Hurtley S. M. Recycling of a secretory granule membrane protein after stimulated secretion. J Cell Sci. 1993 Oct;106(Pt 2):649–655. doi: 10.1242/jcs.106.2.649. [DOI] [PubMed] [Google Scholar]
- Ishii N., Wadsworth W. G., Stern B. D., Culotti J. G., Hedgecock E. M. UNC-6, a laminin-related protein, guides cell and pioneer axon migrations in C. elegans. Neuron. 1992 Nov;9(5):873–881. doi: 10.1016/0896-6273(92)90240-e. [DOI] [PubMed] [Google Scholar]
- Jiang Y. P., Wang H., D'Eustachio P., Musacchio J. M., Schlessinger J., Sap J. Cloning and characterization of R-PTP-kappa, a new member of the receptor protein tyrosine phosphatase family with a proteolytically cleaved cellular adhesion molecule-like extracellular region. Mol Cell Biol. 1993 May;13(5):2942–2951. doi: 10.1128/mcb.13.5.2942. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Joyner A. L., Skarnes W. C., Rossant J. Production of a mutation in mouse En-2 gene by homologous recombination in embryonic stem cells. Nature. 1989 Mar 9;338(6211):153–156. doi: 10.1038/338153a0. [DOI] [PubMed] [Google Scholar]
- Kennedy T. E., Serafini T., de la Torre J. R., Tessier-Lavigne M. Netrins are diffusible chemotropic factors for commissural axons in the embryonic spinal cord. Cell. 1994 Aug 12;78(3):425–435. doi: 10.1016/0092-8674(94)90421-9. [DOI] [PubMed] [Google Scholar]
- Kerr W. G., Nolan G. P., Serafini A. T., Herzenberg L. A. Transcriptionally defective retroviruses containing lacZ for the in situ detection of endogenous genes and developmentally regulated chromatin. Cold Spring Harb Symp Quant Biol. 1989;54(Pt 2):767–776. doi: 10.1101/sqb.1989.054.01.090. [DOI] [PubMed] [Google Scholar]
- Longo F. M., Martignetti J. A., Le Beau J. M., Zhang J. S., Barnes J. P., Brosius J. Leukocyte common antigen-related receptor-linked tyrosine phosphatase. Regulation of mRNA expression. J Biol Chem. 1993 Dec 15;268(35):26503–26511. [PubMed] [Google Scholar]
- Lowell C. A., Soriano P., Varmus H. E. Functional overlap in the src gene family: inactivation of hck and fgr impairs natural immunity. Genes Dev. 1994 Feb 15;8(4):387–398. doi: 10.1101/gad.8.4.387. [DOI] [PubMed] [Google Scholar]
- Mountford P., Zevnik B., Düwel A., Nichols J., Li M., Dani C., Robertson M., Chambers I., Smith A. Dicistronic targeting constructs: reporters and modifiers of mammalian gene expression. Proc Natl Acad Sci U S A. 1994 May 10;91(10):4303–4307. doi: 10.1073/pnas.91.10.4303. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nieto M. A., Gilardi-Hebenstreit P., Charnay P., Wilkinson D. G. A receptor protein tyrosine kinase implicated in the segmental patterning of the hindbrain and mesoderm. Development. 1992 Dec;116(4):1137–1150. doi: 10.1242/dev.116.4.1137. [DOI] [PubMed] [Google Scholar]
- Rudnicki M. A., Schnegelsberg P. N., Stead R. H., Braun T., Arnold H. H., Jaenisch R. MyoD or Myf-5 is required for the formation of skeletal muscle. Cell. 1993 Dec 31;75(7):1351–1359. doi: 10.1016/0092-8674(93)90621-v. [DOI] [PubMed] [Google Scholar]
- Saito H. Structural diversity of eukaryotic protein tyrosine phosphatases: functional and evolutionary implications. Semin Cell Biol. 1993 Dec;4(6):379–387. doi: 10.1006/scel.1993.1045. [DOI] [PubMed] [Google Scholar]
- Stein P. L., Vogel H., Soriano P. Combined deficiencies of Src, Fyn, and Yes tyrosine kinases in mutant mice. Genes Dev. 1994 Sep 1;8(17):1999–2007. doi: 10.1101/gad.8.17.1999. [DOI] [PubMed] [Google Scholar]
- Streuli M., Krueger N. X., Hall L. R., Schlossman S. F., Saito H. A new member of the immunoglobulin superfamily that has a cytoplasmic region homologous to the leukocyte common antigen. J Exp Med. 1988 Nov 1;168(5):1523–1530. doi: 10.1084/jem.168.5.1523. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tashiro K., Tada H., Heilker R., Shirozu M., Nakano T., Honjo T. Signal sequence trap: a cloning strategy for secreted proteins and type I membrane proteins. Science. 1993 Jul 30;261(5121):600–603. doi: 10.1126/science.8342023. [DOI] [PubMed] [Google Scholar]
- Yenofsky R. L., Fine M., Pellow J. W. A mutant neomycin phosphotransferase II gene reduces the resistance of transformants to antibiotic selection pressure. Proc Natl Acad Sci U S A. 1990 May;87(9):3435–3439. doi: 10.1073/pnas.87.9.3435. [DOI] [PMC free article] [PubMed] [Google Scholar]