Binary system for regulating transgene expression in mice: targeting int-2 gene expression with yeast GAL4/UAS control elements.

Ornitz, David M.; Moreadith, Randall W.; Leder, Philip

doi:10.1073/pnas.88.3.698

Cited by 179 publications

(118 citation statements)

References 32 publications

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“…We have used a GA L4/UAS bigenic system (Ornitz et al, 1991;Brand and Perrimon, 1993;Wang et al, 1997), which allows for the production of stable transgenic lines to produce large numbers of embryos that express a lethal transgene, to explore the effects of maintaining ectopic Shh activity in the dorsal neural tube as a model of deregulated Hedgehog signaling in the developing CNS. Analysis of bigenic embryos revealed dramatic neural hyperplasia and enhanced proliferative levels at 12.5 dpc.…”

Section: Discussionmentioning

confidence: 99%

“…However, this resulted in a lethal CNS malformation and precluded maintenance of stable lines. Therefore, on the basis of the work of and Brand and Perrimon (1993) in Drosophila and Ornitz et al (1991) in mice, we adapted the GAL4/UAS bigenic system for controlled gene expression in the developing murine CNS. Six lines of transgenic mice were generated in which GAL4 was expressed under control of Wnt-1 regulatory sequences (Wnt-1/GAL4) (Fig.…”

Section: Wnt-1 Domainmentioning

confidence: 99%

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Sonic hedgehogRegulates Proliferation and Inhibits Differentiation of CNS Precursor Cells

et al. 1999

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Activation of the Sonic hedgehog (Shh) signal transduction pathway is essential for normal pattern formation and cellular differentiation in the developing CNS. However, it is also thought to be etiological in primitive neuroectodermal tumors. We adapted GAL4/UAS methodology to ectopically express full-length Shh in the dorsal neural tube of transgenic mouse embryos commencing at 10 d postcoitum (dpc), beyond the period of primary dorsal-ventral pattern formation and floorplate induction. Expression of Shh was maintained until birth, permitting us to investigate effects of ongoing exposure to Shh on CNS precursors in vivo. Proliferative rates of spinal cord precursors were twice that of wild-type littermates at 12.5 dpc. In contrast, at late fetal stages (18.5 dpc), cells that were Shh-responsive but postmitotic were present in persistent structures reminiscent of the ventricular zone germinal matrix. This tissue remained blocked in an undifferentiated state. These results indicate that cellular competence restricts the proliferative response to Shh in vivo and provide evidence that proliferation and differentiation can be regulated separately in precursor cells of the spinal cord. Thus, Hedgehog signaling may contribute to CNS tumorigenesis by directly enhancing proliferation and preventing neural differentiation in selected precursor cells.

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Section: Discussionmentioning

confidence: 99%

Section: Wnt-1 Domainmentioning

confidence: 99%

Sonic hedgehogRegulates Proliferation and Inhibits Differentiation of CNS Precursor Cells

et al. 1999

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“…First, the intermediate plasmid, pN-DID-HA, was generated by trimolecular ligation of EcoRI digested pcDL-SRa296 (Takebe et al, 1988), the 2.4 kb EcoRI-NdeI fragment from the rat ErbB2 cDNA (Bargmann et al, 1986), and the 236 bp NdeI ± EcoRI digested PCR product generated from the rat ErbB2 cDNA using a forward oligonucleotide primer upstream of the unique NdeI site and the reverse primer 5' GAATGAAT TCAGGCGTAATCAGGCACATCGTATGGGTACAGCC-TACGCATCGTATAC. The modi®ed rat ErbB2 cDNA was then subcloned, to generate the plasmid pMMTV-ErbB2DIC, via a trimolecular ligation involving HindIII ± EcoRI digested pMMTV-Sv40-Bssk (pMMTV-GAL4/236-SV40 minus the GAL4/236 gene) (Ornitz et al, 1991) generously provided by Philip Leder, the 530 bp HindIII ± AatII digested PCR product generated from pN-DID-HA using the forward primer with a 5' HindIII linker 5'-CTAAGCTTCAATGAT-CATCATGGAGCT-3' and the reverse primer 5'-GGGGCA-CAAGGTGGACAGGC-3', and the 3' ca. 1.6 kb AatII ± EcoRI fragment from pN-DID-HA.…”

Section: Plasmidsmentioning

confidence: 99%

Expression of dominant-negative ErbB2 in the mammary gland of transgenic mice reveals a role in lobuloalveolar development and lactation

1999

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Overexpression of the receptor tyrosine kinase ErbB2/ HER2/Neu (ErbB2) occurs in 15 ± 40% of human breast cancers. To determine the function of ErbB2 signaling during normal mouse mammary gland development, we expressed a carboxyl-terminal truncated dominant negative allele of ErbB2 (ErbB2DIC) in the developing mouse mammary gland. Despite ErbB2DIC expression within mammary glands of pubescent virgin and pregnant mice, a phenotype was not observed until late in pregnancy. At 1 day post-partum, lactationally active, distended lobuloalveoli failed to form. This phenotype was exaggerated in multiparous females expressing ErbB2DIC. Immunohistochemical staining for ErbB2-DIC revealed a concordance between high levels of ErbB2DIC protein expression and the absence of lactational products within the lumens of ErbB2DIC stained lobuloalveoli. These results demonstrate that ErbB2 signaling is required for proper mammary development and lactation at parturition.

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“…Preliminary experiments show that the DT-A 151 * and R can easily be integrated in the fly genome under the control of other regulatory sequences such as the yeast regulatory sequences (UAS) that respond only to the yeast transcriptional activator GAL4. Hence, these toxins may become useful tools to express the DT-A ts in tissues other than the photoreceptor cells using a binary system (Ornitz et al, 1991) with on the one hand, enhancer detector strains (Bellen et al, 1989;) that express the GAL4 gene instead of the /Sgalactosidase gene, and, on the other hand, the strains that carry the UAS-DT-A 15 . A.…”

Section: Discussionmentioning

confidence: 99%

Isolation of temperature-sensitive diphtheria toxins in yeast and their effects on Drosophila cells

Bellen

D'Evelyn²,

Harvey³

et al. 1992

Trends in Genetics

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We have isolated temperature-sensitive diphtheria toxins (DT-A**) to develop a method that allows temporal impedement of cellular functions. Four DT-A" 5 genes were isolated in a mutagenesis screen using the yeast, Saccharomyces cerevisiae. When expressed in yeast, these DT-A U arrest growth at 18°C but not at 30°C. Three DT-A" were subsequently tested in the R1-R6 photoreceptor cells of transgenic fruit flies, Drosophila melanogaster. The toxins show similar temperature dependence in both organisms, suggesting that they may be useful in a wide range of non-homeothermic species. DNA sequence analysis revealed that three of the four DT-A 15 mutations are novel. Interestingly, the fourth DT-A te carries the same point mutation as the extensively characterized CRM197, an ADP ribosyltransferase-defective form of diphtheria toxin.

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Binary system for regulating transgene expression in mice: targeting int-2 gene expression with yeast GAL4/UAS control elements.

Cited by 179 publications

References 32 publications

Sonic hedgehogRegulates Proliferation and Inhibits Differentiation of CNS Precursor Cells

Sonic hedgehogRegulates Proliferation and Inhibits Differentiation of CNS Precursor Cells

Expression of dominant-negative ErbB2 in the mammary gland of transgenic mice reveals a role in lobuloalveolar development and lactation

Isolation of temperature-sensitive diphtheria toxins in yeast and their effects on Drosophila cells

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