title: Role of RNA splicing in mediating lineage-specific expression of the von Willebrand factor gene in the endothelium.
abstract: We previously demonstrated that the first intron of the human von Willebrand factor (vWF) is required for gene expression in the endothelium of transgenic mice. Based on this finding, we hypothesized that RNA splicing plays a role in mediating vWF expression in the vasculature. To address this question, we used transient transfection assays in human endothelial cells and megakaryocytes with intron-containing and intronless human vWF promoter-luciferase constructs. Next, we generated knockin mice in which LacZ was targeted to the endogenous mouse vWF locus in the absence or presence of the native first intron or heterologous introns from the human β-globin, mouse Down syndrome critical region 1, or hagfish coagulation factor X genes. In both the in vitro assays and the knockin mice, the loss of the first intron of vWF resulted in a significant reduction of reporter gene expression in endothelial cells but not megakaryocytes. This effect was rescued to varying degrees by the introduction of a heterologous intron. Intron-mediated enhancement of expression was mediated at a posttranscriptional level. Together, these findings implicate a role for intronic splicing in mediating lineage-specific expression of vWF in the endothelium.
authors.affiliation: Center for Vascular Biology Research, Department of Medicine, Beth Israel Deaconess Medical Center, Boston, MA 02215, USA.
authors.forename: Lei
authors.initials: L
authors.lastname: Yuan
authors.forename: Lauren
authors.initials: L
authors.lastname: Janes
authors.forename: David
authors.initials: D
authors.lastname: Beeler
authors.forename: Katherine C
authors.initials: KC
authors.lastname: Spokes
authors.forename: Joshua
authors.initials: J
authors.lastname: Smith
authors.forename: Dan
authors.initials: D
authors.lastname: Li
authors.forename: Shou-Ching
authors.initials: SC
authors.lastname: Jaminet
authors.forename: Peter
authors.initials: P
authors.lastname: Oettgen
authors.forename: William C
authors.initials: WC
authors.lastname: Aird
mesh_terms: D000818:Animals; D002417:Cattle; D019070:Cell Lineage / Q000235:genetics*; D004730:Endothelium, Vascular / Q000502:physiology*; D005091:Exons / Q000235:genetics; D055879:Gene Knock-In Techniques; D006487:Hemostasis / Q000502:physiology; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007438:Introns / Q000235:genetics; D007763:Lac Operon; D051379:Mice; D011401:Promoter Regions, Genetic / Q000235:genetics; D012326:RNA Splicing / Q000235:genetics*; D013045:Species Specificity; D014841:von Willebrand Factor / Q000235:genetics*
chemical_list: D014841:von Willebrand Factor