title: Performance of genomic bordering elements at predefined genomic loci.
abstract: Eukaryotic DNA is organized into chromatin domains that regulate gene expression and chromosome behavior. Insulators and/or scaffold-matrix attachment regions (S/MARs) mark the boundaries of these chromatin domains where they delimit enhancing and silencing effects from the outside. By recombinase-mediated cassette exchange (RMCE), we were able to compare these two types of bordering elements at a number of predefined genomic loci. Flanking an expression vector with either S/MARs or two copies of the non-S/MAR chicken hypersensitive site 4 insulator demonstrates that while these borders confer related expression characteristics at most loci, their effect on chromatin organization is clearly distinct. Our results suggest that the activity of bordering elements is most pronounced for the abundant class of loci with a low but negligible expression potential in the case of highly expressed sites. By the RMCE procedure, we demonstrate that expression parameters are not due to a potential targeting action of bordering elements, in the sense that a linked transgene is directed into a special class of loci. Instead, we can relate the observed transcriptional augmentation phenomena to their function as genomic insulators.
authors.affiliation: German Research Centre for Biotechnology (GBF), RDIF/Epigenetic Regulation, Mascheroder Weg 1, 38124 Braunschweig, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Goetze
authors.forename: Alexandra
authors.initials: A
authors.lastname: Baer
authors.forename: Silke
authors.initials: S
authors.lastname: Winkelmann
authors.forename: Kristina
authors.initials: K
authors.lastname: Nehlsen
authors.forename: Jost
authors.initials: J
authors.lastname: Seibler
authors.forename: Karin
authors.initials: K
authors.lastname: Maass
authors.forename: Jürgen
authors.initials: J
authors.lastname: Bode
mesh_terms: D000818:Animals; D002460:Cell Line; D002645:Chickens / Q000235:genetics; D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D021901:DNA, Intergenic / Q000235:genetics / Q000502:physiology; D005786:Gene Expression Regulation; D017930:Genes, Reporter / Q000235:genetics; D016678:Genome; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D038101:Insulator Elements / Q000235:genetics / Q000502:physiology*; D045170:Matrix Attachment Regions / Q000235:genetics*; D051379:Mice; D045522:Recombinases / Q000235:genetics / Q000502:physiology; D011995:Recombination, Genetic / Q000235:genetics; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
chemical_list: D002843:Chromatin; D021901:DNA, Intergenic; D045522:Recombinases; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase
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