Functional analysis of secreted and transmembrane proteins critical to mouse development

Mitchell, Kevin J.; Pinson, Kathy; Kelly, Olivia G.; Brennan, Jane; Zupicich, Joel; Scherz, Paul; Leighton, Philip A.; Goodrich, Lisa V.; Lu, Xiaowei; Avery, Brian J.; Täte, P.; Dill, Kariena K.; Pangilinan, Edivinia; Wakenight, Paul; Tessier‐Lavigne, Marc; Skarnes, William C.

doi:10.1038/90074

Cited by 380 publications

(356 citation statements)

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“…The trapped genes were distributed on all chromosomes except the Y, suggesting that a large number of genes across the genome could be accessed by this vector. Some loci seem to be more readily trapped than others, as also found by other large-scale gene trapping efforts using different vectors [19][20][21][22] , and 40 genes were trapped more than once. This phenomenon may be due to recombination hot spots, more permissive integration sites available in larger genes or a limited pool of genes that could be trapped in the genome.…”

Section: Cloning and Analysis Of Trapped Transcriptssupporting

confidence: 68%

“…But all these screens are confined to ES cells or their differentiated derivatives and rely on reporter expression. Sequence-driven screens based on the identity of trapped genes [19][20][21][22]30 are intrinsically biased towards highly annotated genes, and limited functional cues can be deduced from the trapped sequences representing ESTs or new genes. With the gene-trap array, virtually any cell type can now be screened for differentially expressed genes.…”

Section: Discussionmentioning

confidence: 99%

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Identification and validation of PDGF transcriptional targets by microarray-coupled gene-trap mutagenesis

et al. 2004

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We developed a versatile, high-throughput genetic screening strategy by coupling gene mutagenesis and expression profiling technologies. Using a retroviral gene-trap vector optimized for efficient mutagenesis and cloning, we randomly disrupted genes in mouse embryonic stem (ES) cells and amplified them to construct a cDNA microarray. With this gene-trap array, we show that transcriptional target genes of platelet-derived growth factor (PDGF) can be efficiently and reliably identified in physiologically relevant cells and are immediately accessible to genetic studies to determine their in vivo roles and relative contributions to PDGF-regulated developmental processes. The same platform can be used to search for genes of specific biological relevance in a broad array of experimental settings, providing a fast track from gene identification to functional validation.

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Section: Cloning and Analysis Of Trapped Transcriptssupporting

confidence: 68%

Section: Discussionmentioning

confidence: 99%

Identification and validation of PDGF transcriptional targets by microarray-coupled gene-trap mutagenesis

et al. 2004

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“…Mice carrying a gene-trap insertion in the Neo1 gene have been constructed as described previously (Leighton et al, 2001;Mitchell et al, 2001), and homozygous mutants were reported to display perinatal lethality (Srinivasan et al, 2003), but this allele is not well characterized. We refer to the allele here as Neo1 Gt(KST265)Byg (abbreviated Neo1 Gt ).…”

Section: The Neo1 Gt Allele Is Hypomorphic and Results In Incompletelmentioning

confidence: 99%

“…Mice carrying a secretory gene-trap vector insertion into intron 7 of the Neo1 gene were constructed previously (Leighton et al, 2001;Mitchell et al, 2001). The embryonic stem (ES) cell line that carried the secretory gene trap from which this mouse line was developed was designated KST265, and we therefore refer to the allele as Neo1 Gt(KST265)Byg (abbreviated Neo1 Gt ).…”

Section: Micementioning

confidence: 99%

“…The insertion is predicted to produce a mutant transcript that encodes a transmembrane protein consisting of the four Ig repeats of neogenin fused in-frame with a vector-encoded transmembrane domain and a cytoplasmic domain consisting of ␤-geo (␤-galactosidase fused to neomycin phosphotransferase). Previous studies suggest that such fusion proteins are targeted to intracellular compartments (Leighton et al, 2001;Mitchell et al, 2001).All mice and embryos were of a largely C57BL/6 background and were generated from intercrosses of Neo1 ϩ/Gt animals. Noon of the plug date was designated E0.5.…”

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Neogenin Regulates Skeletal Myofiber Size and Focal Adhesion Kinase and Extracellular Signal-regulated Kinase Activities In Vivo and In Vitro

Bae

Yang

Jiang

et al. 2009

MBoC

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A variety of signaling pathways participate in the development of skeletal muscle, but the extracellular cues that regulate such pathways in myofiber formation are not well understood. Neogenin is a receptor for ligands of the netrin and repulsive guidance molecule (RGM) families involved in axon guidance. We reported previously that neogenin promoted myotube formation by C2C12 myoblasts in vitro and that the related protein Cdo (also Cdon) was a potential neogenin coreceptor in myoblasts. We report here that mice homozygous for a gene-trap mutation in the Neo1 locus (encoding neogenin) develop myotomes normally but have small myofibers at embryonic day 18.5 and at 3 wk of age. Similarly, cultured myoblasts derived from such animals form smaller myotubes with fewer nuclei than myoblasts from control animals. These in vivo and in vitro defects are associated with low levels of the activated forms of focal adhesion kinase (FAK) and extracellular signal-regulated kinase (ERK), both known to be involved in myotube formation, and inefficient expression of certain muscle-specific proteins. Recombinant netrin-2 activates FAK and ERK in cultured myoblasts in a neogenin-and Cdo-dependent manner, whereas recombinant RGMc displays lesser ability to activate these kinases. Together, netrin-neogenin signaling is an important extracellular cue in regulation of myogenic differentiation and myofiber size. INTRODUCTIONSkeletal muscle is the most abundant tissue, by mass, in the vertebrate body. Muscles of the trunk and limbs arise from the somites, with myogenic progenitor cells derived from the dorsal region of the maturing somite, the dermomyotome (Tajbakhsh and Buckingham, 2000;Pownall et al., 2002;Charge and Rudnicki, 2004). In response to signals from the adjacent notochord, neural tube, and surface ectoderm, some dermomyotomal progenitors become committed to the muscle lineage and form the myotome, a set of differentiated muscle cells that underlies the dermomyotome. Subsequent embryonic, fetal, and postnatal stages of myogenesis are thought to involve additional muscle progenitors that migrate from the dermomyotome and ultimately establish the trunk and limb musculature, as well as satellite cells, adult muscle precursor cells (Gros et al., 2005;Kassar-Duchossoy et al., 2005;Relaix et al., 2005). Somitic progenitor cells are specified to become muscle lineage-committed myoblasts through the action of the myogenic basic helix-loop-helix transcription factors Myf5, MRF4, and MyoD, whereas differentiation of myoblasts is regulated by myogenin, MyoD, and MRF4 (Tajbakhsh, 2005). These and other transcription factors coordinate the process of differentiation, including cell cycle withdrawal, expression of muscle-specific proteins, cellular elongation, and fusion into multinucleated myofibers. Although transcriptional regulation is at the core of myogenesis, the formation and growth of myofibers is also controlled by a variety of signaling ligands and their receptors, including insulin-like growth factor-1, fibroblast growth fac...

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Mouse mutagenesis and gene function

Kühn¹,

Wurst

2005

Encyclopedia of Genetics, Genomics, Proteomics and Bioinformatics

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With the completion of the mouse and human genome sequences, a major challenge is the functional characterization of every gene within the mammalian genome. The mouse offers many advantages for the use of genetics to the study of human biology and disease. A variety of mouse mutagenesis technologies, either gene‐ or phenotype‐driven, are used as systematic approaches. The availability of the mouse genome supports gene‐driven approaches such as gene‐trap and targeted mutagenesis in embryonic stem (ES) cells, allowing an efficiency and precision of gene disruption unmatched among other mammals. Furthermore, chemical and transposon mutagenesis of the mouse genome allows to perform phenotype‐driven screens for the unbiased identification of phenotype–genotype correlations. These technologies already resulted in a collection of several thousand mutants. In this article on mouse mutagenesis and gene function, we present a comprehensive review of gene‐ and phenotype‐driven mouse mutagenesis strategies. Besides a summary of the basic principles of each approach, we emphasize on the latest and future developments of mouse mutagenesis.

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Functional analysis of secreted and transmembrane proteins critical to mouse development

Cited by 380 publications

References 53 publications

Identification and validation of PDGF transcriptional targets by microarray-coupled gene-trap mutagenesis

Identification and validation of PDGF transcriptional targets by microarray-coupled gene-trap mutagenesis

Neogenin Regulates Skeletal Myofiber Size and Focal Adhesion Kinase and Extracellular Signal-regulated Kinase Activities In Vivo and In Vitro

Mouse mutagenesis and gene function

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