Use of large-scale expression cloning screens in the Xenopus laevis tadpole to identify gene function.

We have conducted an expression cloning screen of approximately 50, 000 cDNAs from a tadpole stage Xenopus laevis cDNA library to functionally identify genes affecting a wide range of cellular and developmental processes. Fifty-seven cDNAs were isolated for their ability to alter gross tadpole morphology or the expression patterns of tissue-specific markers. Thirty-seven of the cDNAs have not been previously described for Xenopus, and 15 of these show little or no similarity to sequences in the NCBI database. The screen and the identified genes are presented in this paper to demonstrate the power, ease, speed, and flexibility of expression cloning in the X. laevis embryo. Future screens such as this one can be done on a larger scale and will complement the sequence-based screens and genome-sequencing projects which are producing a large body of novel genes without ascribed functions.

[1]  Haigen Huang,et al.  Stimulation of erythropoiesis by inhibiting a new hematopoietic death receptor in transgenic zebrafish , 2000, Nature Cell Biology.

[2]  T. Pawson,et al.  A mammalian PAR-3–PAR-6 complex implicated in Cdc42/Rac1 and aPKC signalling and cell polarity , 2000, Nature Cell Biology.

[3]  M. Bronner‐Fraser,et al.  Snail-related transcriptional repressors are required in Xenopus for both the induction of the neural crest and its subsequent migration. , 2000, Developmental biology.

[4]  K. Mizuseki,et al.  Requirement of Sox2-mediated signaling for differentiation of early Xenopus neuroectoderm. , 2000, Development.

[5]  W. McGinnis,et al.  Developmental patterning genes and their conserved functions: from model organisms to humans. , 2000, Molecular genetics and metabolism.

[6]  R. Beddington,et al.  Wnt signaling in Xenopus embryos inhibits bmp4 expression and activates neural development. , 1999, Genes & development.

[7]  I. Blitz,et al.  Involvement of the small GTPases XRhoA and XRnd1 in cell adhesion and head formation in early Xenopus development. , 1999, Development.

[8]  H. Friess,et al.  Reduced expression of the metastasis suppressor gene KAI1 in advanced colon cancer and its metastases. , 1999, Surgery.

[9]  K. Takeshima,et al.  The early expression control of Xepsin by nonaxial and planar posteriorizing signals in Xenopus epidermis. , 1999, Developmental biology.

[10]  P. D. Vize,et al.  Synergism between Pax-8 and lim-1 in embryonic kidney development. , 1999, Developmental biology.

[11]  M. Beanan,et al.  Inhibitory patterning of the anterior neural plate in Xenopus by homeodomain factors Dlx3 and Msx1. , 1999, Developmental biology.

[12]  M. Kai,et al.  Molecular characterization of the type 2 phosphatidic acid phosphatase. , 1999, Chemistry and physics of lipids.

[13]  R. Harland,et al.  XBF-2 is a transcriptional repressor that converts ectoderm into neural tissue. , 1998, Development.

[14]  L. Zon,et al.  Cloning of Mix-related homeodomain proteins using fast retrieval of gel shift activities, (FROGS), a technique for the isolation of DNA-binding proteins. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[15]  G. Salvesen,et al.  Properties of the caspases. , 1998, Biochimica et biophysica acta.

[16]  J. Marden,et al.  FGF-mediated mesoderm induction involves the Src-family kinase Laloo , 1998, Nature.

[17]  M. Kirschner,et al.  Geminin, a neuralizing molecule that demarcates the future neural plate at the onset of gastrulation. , 1998, Development.

[18]  J. Barrett,et al.  Frequent downregulation of the KAI1(CD82) metastasis suppressor protein in human cancer cell lines , 1998, Oncogene.

[19]  M. Kirschner,et al.  Geminin, an Inhibitor of DNA Replication, Is Degraded during Mitosis , 1998, Cell.

[20]  S. Chen,et al.  CIDE, a novel family of cell death activators with homology to the 45 kDa subunit of the DNA fragmentation factor , 1998, The EMBO journal.

[21]  Thomas M. Jessell,et al.  Molecular and cellular approaches to neural development , 1998 .

[22]  R. Harland,et al.  The Xenopus dorsalizing factor Gremlin identifies a novel family of secreted proteins that antagonize BMP activities. , 1998, Molecular cell.

[23]  K. Mizuseki,et al.  Xenopus Zic-related-1 and Sox-2, two factors induced by chordin, have distinct activities in the initiation of neural induction. , 1998, Development.

[24]  C. Niehrs,et al.  Dickkopf-1 is a member of a new family of secreted proteins and functions in head induction , 1998, Nature.

[25]  M. Kirschner,et al.  Sizzled: a secreted Xwnt8 antagonist expressed in the ventral marginal zone of Xenopus embryos. , 1997, Development.

[26]  Ken W. Y. Cho,et al.  The Xenopus homeobox gene twin mediates Wnt induction of goosecoid in establishment of Spemann's organizer. , 1997, Development.

[27]  J. Gautier,et al.  A developmental timer that regulates apoptosis at the onset of gastrulation , 1997, Mechanisms of Development.

[28]  N. Dahmane,et al.  Activation of the transcription factor Gli1 and the Sonic hedgehog signalling pathway in skin tumours , 1997, Nature.

[29]  Jay X. Tang,et al.  Caspase-3-generated fragment of gelsolin: effector of morphological change in apoptosis. , 1997, Science.

[30]  C. Niehrs,et al.  Head induction by simultaneous repression of Bmp and Wnt signalling in Xenopus , 1997, Nature.

[31]  J. Newport,et al.  Developmentally regulated activation of apoptosis early in Xenopus gastrulation results in cyclin A degradation during interphase of the cell cycle. , 1997, Development.

[32]  N. Ueno,et al.  Xenopus msx1 mediates epidermal induction and neural inhibition by BMP4. , 1997, Development.

[33]  M. Kirschner,et al.  FUNCTIONAL GENOMICS: Expression Cloning in the Test Tube , 1997, Science.

[34]  A. Lewellyn,et al.  Ionizing radiation induces apoptosis and elevates cyclin A1-Cdk2 activity before but not after the midblastula transition in Xenopus. , 1997, Molecular biology of the cell.

[35]  M. Kirschner,et al.  Systematic identification of mitotic phosphoproteins , 1997, Current Biology.

[36]  S. Levy,et al.  The tetraspanin superfamily: molecular facilitators , 1997, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[37]  N. Ueno,et al.  Mesoderm induction by BMP-4 and -7 heterodimers. , 1997, Biochemical and biophysical research communications.

[38]  M. Kirschner,et al.  Expression cloning of a Xenopus T-related gene (Xombi) involved in mesodermal patterning and blastopore lip formation. , 1996, Development.

[39]  J. Baker,et al.  A novel mesoderm inducer, Madr2, functions in the activin signal transduction pathway. , 1996, Genes & development.

[40]  R. Harland,et al.  A nodal-related gene defines a physical and functional domain within the Spemann organizer , 1995, Cell.

[41]  P. Lemaire,et al.  Expression cloning of Siamois, a xenopus homeobox gene expressed in dorsal-vegetal cells of blastulae and able to induce a complete secondary axis , 1995, Cell.

[42]  I. Blitz,et al.  Anterior neurectoderm is progressively induced during gastrulation: the role of the Xenopus homeobox gene orthodenticle. , 1995, Development.

[43]  E. Boncinelli,et al.  The Xenopus homologue of Otx2 is a maternal homeobox gene that demarcates and specifies anterior body regions. , 1995, Development.

[44]  R. Mayor,et al.  Induction of the prospective neural crest of Xenopus. , 1995, Development.

[45]  N. Ueno,et al.  A truncated bone morphogenetic protein receptor affects dorsal-ventral patterning in the early Xenopus embryo. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[46]  R. Harland.  Neural induction in Xenopus. , 1994, Current opinion in genetics & development.

[47]  H. Weintraub,et al.  Expression of achaete-scute homolog 3 in Xenopus embryos converts ectodermal cells to a neural fate. , 1994, Genes & development.

[48]  T. Lints,et al.  XNkx-2.5, a Xenopus gene related to Nkx-2.5 and tinman: evidence for a conserved role in cardiac development. , 1994, Developmental biology.

[49]  R. Harland,et al.  XFKH2, a Xenopus HNF-3 alpha homologue, exhibits both activin-inducible and autonomous phases of expression in early embryos. , 1993, Developmental biology.

[50]  William C. Smith,et al.  Expression cloning of noggin, a new dorsalizing factor localized to the Spemann organizer in Xenopus embryos , 1992, Cell.

[51]  D. Melton,et al.  Involvement of p21ras in Xenopus mesoderm induction , 1992, Nature.

[52]  L. Zon,et al.  Expression of GATA-binding proteins during embryonic development in Xenopus laevis. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[53]  R. Harland,et al.  Injected Xwnt-8 RNA acts early in Xenopus embryos to promote formation of a vegetal dorsalizing center , 1991, Cell.

[54]  Douglas A. Melton,et al.  Injected Wnt RNA induces a complete body axis in Xenopus embryos , 1991, Cell.

[55]  D. Wilkinson,et al.  Conserved segmental expression of Krox-20 in the vertebrate hindbrain and its relationship to lineage restriction. , 1991, Development (Cambridge, England). Supplement.

[56]  R. Moon,et al.  Xwnt-8, a Xenopus Wnt-1/int-1-related gene responsive to mesoderm-inducing growth factors, may play a role in ventral mesodermal patterning during embryogenesis. , 1991, Development.

[57]  M. Somerman,et al.  Fibulin is an extracellular matrix and plasma glycoprotein with repeated domain structure , 1990, The Journal of cell biology.

[58]  D. Lowy,et al.  Identification of small clusters of divergent amino acids that mediate the opposing effects of ras and Krev-1. , 1990, Science.

[59]  P. Good,et al.  A developmentally regulated, nervous system-specific gene in Xenopus encodes a putative RNA-binding protein. , 1990, The New biologist.

[60]  J. Gurdon,et al.  MyoD expression in the forming somites is an early response to mesoderm induction in Xenopus embryos. , 1989, The EMBO journal.

[61]  Andrew P. McMahon,et al.  Ectopic expression of the proto-oncogene int-1 in Xenopus embryos leads to duplication of the embryonic axis , 1989, Cell.

[62]  J. Gurdon,et al.  A homeobox-containing marker of posterior neural differentiation shows the importance of predetermination in neural induction , 1987, Cell.

[63]  S. Moody,et al.  Fates of the blastomeres of the 16-cell stage Xenopus embryo. , 1987, Developmental biology.

[64]  J M Slack,et al.  Fate map for the 32-cell stage of Xenopus laevis. , 1987, Development.

[65]  J. Faber,et al.  Normal Table of Xenopus Laevis (Daudin) , 1958 .

[66]  R. Harland,et al.  Early development of Xenopus laevis : a laboratory manual , 2000 .

[67]  J. Nelson,et al.  Prostate cancer metastasis-suppressor genes: a current perspective. , 1998, In vivo.

[68]  D. Ron.  TLS-CHOP and the role of RNA-binding proteins in oncogenic transformation. , 1997, Current topics in microbiology and immunology.

[69]  L. Zon,et al.  Small pool expression screening: identification of genes involved in cell cycle control, apoptosis, and early development. , 1997, Methods in enzymology.

[70]  J. Gerhart,et al.  Formation and function of Spemann's organizer. , 1997, Annual review of cell and developmental biology.

[71]  T. Fischer,et al.  Cytoskeletal Interactions of Raplb in Platelets , 1993 .

[72]  M. McCormick.  [33] Sib selection , 1987 .