0000000000588004

AUTHOR

Paola Di Simone

showing 2 related works from this author

The Sea Urchin sns Insulator Blocks CMV Enhancer following Integration in Human Cells

2001

Insulators are a new class of genetic elements that attenuate enhancer function directionally. Previously, we characterized in sea urchin a 265-bp-long insulator, termed sns. To test insulator activity following stable integration in human cells, we placed sns between the CMV enhancer and a tk promoter up-stream of a GFP transgene of plasmid or retroviral vectors. In contrast to controls, cells transfected or transduced with insulated constructs displayed a barely detectable fluorescence. Southern blot and PCR ruled out vector rearrangement following integration into host DNA; RNase protection confirmed the enhancer blocking activity. Finally, we demonstrate that two cis-acting sequences, p…

animal structuresSea UrchinVirus IntegrationTransgeneMolecular Sequence DataBiophysicsCytomegalovirusSettore BIO/11 - Biologia MolecolareSimian virus 40BiologyTransfectionPolymerase Chain ReactionBiochemistrySodium ChannelsNAV1.8 Voltage-Gated Sodium ChannelPlasmidTumor Cells CulturedAnimalsHumansEnhancer trapDNA Polymerase Chain ReactionEnhancerBinding Sites; DNA Polymerase Chain Reaction; Recombinant Proteins; Sea Urchins;Tumor Cells Cultured; Enhancer Elements Genetic; Virus Integration;Molecular BiologyVirus IntegrationSouthern blotBinding SitesBase SequenceBinding SiteCell BiologyTransfectionRecombinant ProteinMolecular biologyRecombinant ProteinsChromatinSettore BIO/18 - GeneticaEnhancer Elements GeneticSea UrchinsDNA ViralBiochemical and Biophysical Research Communications
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Functional characterization of the enhancer blocking element of the sea urchin early histone gene cluster reveals insulator properties and three esse…

2000

Insulator elements can be functionally identified by their ability to shield promoters from regulators in a position-dependent manner or their ability to protect adjacent transgenes from position effects. We have previously reported the identification of a 265 bp sns DNA fragment at the 3' end of the sea urchin H2A early histone gene that blocked expression of a reporter gene in transgenic embryos when placed between the enhancer and the promoter. Here we show that sns interferes with enhancer-promoter interaction in a directional manner. When sns is placed between the H2A modulator and the inducible tet operator, the modulator is barred from interaction with the basal promoter. However, th…

animal structuresenhancer blockingMolecular Sequence DataDNA FootprintingSettore BIO/11 - Biologia MolecolareBiologyRegulatory Sequences Nucleic AcidinsulatorBinding CompetitiveHistonesStructural BiologyTranscription (biology)Gene clustermicroinjectionAnimalsDeoxyribonuclease IH2A enhancerGene SilencingTransgenesEnhancerDownstream EnhancerPromoter Regions GeneticMolecular BiologyTranscription factorRepetitive Sequences Nucleic AcidSequence DeletionReporter geneBase SequenceActivator (genetics)PromoterDNAhistone genesMolecular biologyCell biologyDNA-Binding ProteinsEnhancer Elements GeneticMultigene FamilySea UrchinsProtein Binding
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