Characterization of a gene trap insertion into a novel gene, cordon-bleu, expressed in axial structures of the gastrulating mouse embryo.

We have used a gene trap (GT) vector and embryonic stem (ES) cell chimeras to screen for insertions of the lacZ reporter gene into transcription units that are spatially and temporally regulated during early mouse embryogenesis. GT vectors which can act as both a reporter and a mutagen have been previously used to isolate new genes that are essential for mouse development. In this paper we describe a GT insertion which displays a very restricted pattern of expression in the gastrulating embryo. beta-Galactosidase activity was first detected at 7.5 days post-coitum (E7.5) in the node region of the embryo and extended to the midline structures at E8.0. At E9.5 expression was restricted to the floor plate, the notochord, the roof of the gut, and the liver anlage. Expression appeared in the somites at E10.0 and later became more widespread. We used rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR) to clone a partial 360 base pair (bp) cDNA representing an endogenous sequence and containing an open reading frame (ORF) fused in frame to the lacZ reporter gene. The sequence showed no homology to any known protein or protein domain. An overlapping 1,200 bp fragment from a wild-type cDNA library was cloned and it detected the same pattern of expression as the reporter gene in E7.5, E8.5, and E9.5 wild-type embryos. It hybridized to a 5.4 kb lacZ fusion transcript and to an endogenous transcript of 6.5 kb. The gene was mapped to chromosome 11 and was named cordon-bleu (cobl). No phenotype was detected in mice homozygous for the insertion. However, the insertion may not cause a complete disruption of the gene function. The pattern of expression of cobl is very similar to that of hepatic nuclear factor 3 beta (HNF3 beta) and sonic hedgehog (Shh), both of which are involved in axial patterning. Therefore, the product of the cobl gene may also prove to be an important component of the genetic pathway regulating vertebrate axis formation.

[1]  F. Conlon,et al.  Effects of the TWis mutation on notochord formation and mesodermal patterning , 1995, Mechanisms of Development.

[2]  A. Joyner,et al.  A large-scale gene-trap screen for insertional mutations in developmentally regulated genes in mice. , 1995, Genetics.

[3]  T. Magnuson,et al.  The eed mutation disrupts anterior mesoderm production in mice. , 1995, Development.

[4]  M. Tessier-Lavigne,et al.  Patterning of mammalian somites by surface ectoderm and notochord: Evidence for sclerotome induction by a hedgehog homolog , 1994, Cell.

[5]  C. Tabin,et al.  Ectopic expression of Sonic hedgehog alters dorsal-ventral patterning of somites , 1994, Cell.

[6]  Thomas M. Jessell,et al.  The winged-helix transcription factor HNF-3β is required for notochord development in the mouse embryo , 1994, Cell.

[7]  J. Rossant,et al.  HNF-3β is essential for node and notochord formation in mouse development , 1994, Cell.

[8]  F. Conlon,et al.  A primary requirement for nodal in the formation and maintenance of the primitive streak in the mouse. , 1994, Development.

[9]  C. Théry,et al.  Of mice and frogs. , 1994, Trends in genetics : TIG.

[10]  R. Beddington.  Induction of a second neural axis by the mouse node. , 1994, Development.

[11]  C. Nüsslein-Volhard,et al.  Large-scale mutagenesis in the zebrafish: in search of genes controlling development in a vertebrate , 1994, Current Biology.

[12]  T. Jessell,et al.  Floor plate and motor neuron induction by vhh-1, a vertebrate homolog of hedgehog expressed by the notochord , 1994, Cell.

[13]  B. Hogan,et al.  HNF-3β as a regulator of floor plate development , 1994, Cell.

[14]  C. Tabin,et al.  Sonic hedgehog mediates the polarizing activity of the ZPA , 1993, Cell.

[15]  Andrew P. McMahon,et al.  Sonic hedgehog, a member of a family of putative signaling molecules, is implicated in the regulation of CNS polarity , 1993, Cell.

[16]  P. Gruss,et al.  Altered Pax gene expression in murine notochord mutants: the notochord is required to initiate and maintain ventral identity in the somite , 1993, Mechanisms of Development.

[17]  J. Rossant,et al.  The formation and maintenance of the definitive endoderm lineage in the mouse: involvement of HNF3/forkhead proteins. , 1993, Development.

[18]  J. Darnell,et al.  Sequential expression of HNF-3β and HNF-3α by embryonic organizing centers: the dorsal lip/node, notochord and floor plate , 1993, Mechanisms of Development.

[19]  R. Balling,et al.  A role for Pax-1 as a mediator of notochordal signals during the dorsoventral specification of vertebrae. , 1993, Development.

[20]  J. Roder,et al.  Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[21]  T. Jessell,et al.  Ectopic neural expression of a floor plate marker in frog embryos injected with the midline transcription factor Pintallavis. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[22]  T. Magnuson,et al.  Genetic control of gastrulation in the mouse. , 1993, Current opinion in genetics & development.

[23]  O. Pourquié,et al.  Control of dorsoventral patterning of somitic derivatives by notochord and floor plate. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[24]  B. Hogan,et al.  Differential expression of multiple fork head related genes during gastrulation and axial pattern formation in the mouse embryo. , 1993, Development.

[25]  A. Joyner,et al.  A mouse model of Greig cephalo–polysyndactyly syndrome: the extra–toesJ mutation contains an intragenic deletion of the Gli3 gene , 1993, Nature Genetics.

[26]  Linda Lowe,et al.  Nodal is a novel TGF-β-like gene expressed in the mouse node during gastrulation , 1993, Nature.

[27]  J. Rossant,et al.  Exogenous retinoic acid rapidly induces anterior ectopic expression of murine Hox-2 genes in vivo. , 1992, Development.

[28]  Ken W. Y. Cho,et al.  Gastrulation in the mouse: The role of the homeobox gene goosecoid , 1992, Cell.

[29]  W. C. Forrester,et al.  A gene trap approach in mouse embryonic stem cells: the lacZ reported is activated by splicing, reflects endogenous gene expression, and is mutagenic in mice. , 1992, Genes & development.

[30]  A. Joyner,et al.  A targeted mutation reveals a role for N-myc in branching morphogenesis in the embryonic mouse lung. , 1992, Genes & development.

[31]  Ken W. Y. Cho,et al.  Molecular nature of Spemann's organizer: the role of the Xenopus homeobox gene goosecoid , 1991, Cell.

[32]  R. Pedersen,et al.  Clonal analysis of epiblast fate during germ layer formation in the mouse embryo. , 1991, Development.

[33]  B. Herrmann,et al.  Expression pattern of the Brachyury gene in whole-mount TWis/TWis mutant embryos. , 1991, Development.

[34]  Philippe Soriano,et al.  Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice. , 1991, Genes & development.

[35]  C. Stern,et al.  Fate mapping and cell lineage analysis of Hensen's node in the chick embryo. , 1991, Development.

[36]  T. Jessell,et al.  Mesodermal control of neural cell identity: floor plate induction by the notochord. , 1990, Science.

[37]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[38]  H. V. Melchner,et al.  Isolation of cellular promoters by using a retrovirus promoter trap. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[39]  D. Wilkinson,et al.  Expression pattern of the mouse T gene and its role in mesoderm formation , 1990, Nature.

[40]  A. Poustka,et al.  Cloning of the T gene required in mesoderm formation in the mouse , 1990, Nature.

[41]  M. Capecchi,et al.  Altering the genome by homologous recombination. , 1989, Science.

[42]  A. Joyner,et al.  Mouse embryonic stem cells and reporter constructs to detect developmentally regulated genes. , 1989, Science.

[43]  M. Frohman,et al.  Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[44]  D. Lipman,et al.  Improved tools for biological sequence comparison. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[45]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[46]  T. Doetschman,et al.  Transgenesis by means of blastocyst-derived embryonic stem cell lines. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[47]  R Kemler,et al.  The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. , 1985, Journal of embryology and experimental morphology.

[48]  A. Bradley,et al.  Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines , 1984, Nature.

[49]  A. Feinberg,et al.  A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. , 1983, Analytical biochemistry.

[50]  G. Martin,et al.  Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[51]  M. Kaufman,et al.  Establishment in culture of pluripotential cells from mouse embryos , 1981, Nature.

[52]  C. Nüsslein-Volhard,et al.  Mutations affecting segment number and polarity in Drosophila , 1980, Nature.

[53]  P. Meneely,et al.  Lethals, steriles and deficiencies in a region of the X chromosome of Caenorhabditis elegans. , 1979, Genetics.

[54]  J. Smith,et al.  Control of vertebrate gastrulation: inducing signals and responding genes. , 1993, Current opinion in genetics & development.

[55]  T. Jessell,et al.  Midline cells and the organization of the vertebrate neuraxis. , 1993, Current opinion in genetics & development.

[56]  B. Herrmann,et al.  Detection of messenger RNA by in situ hybridization to postimplantation embryo whole mounts. , 1993, Methods in enzymology.

[57]  E. Rinchik.  Chemical mutagenesis and fine-structure functional analysis of the mouse genome. , 1991, Trends in genetics : TIG.

[58]  T. Jessell,et al.  Floor plate-derived signals and the control of neural cell pattern in vertebrates. , 1990, Harvey lectures.

[59]  W. Dove,et al.  Saturation germ line mutagenesis of the murine t region including a lethal allele at the quaking locus. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[60]  T. Magnuson,et al.  The albino deletion complex and early postimplantation survival in the mouse. , 1988, Development.

[61]  C. H. Waddington,et al.  Induction by the Primitive Streak and its Derivatives in the Chick , 1933 .