Targeting gene expression to endothelium in transgenic animals: a comparison of the human ICAM‐2, PECAM‐1 and endoglin promoters

Cowan, Peter J.; Shinkel, Trixie A.; Fisicaro, Nella; Godwin, James; Bernabéu, Carmelo; Almendro, Nuria; Rı́us, Carlos; Lonie, Andrew; Nottle, Mark B.; Wigley, Peter L.; Paizis, Kathy; Pearse, Martin J.; d’Apice, Anthony J.F.

doi:10.1034/j.1399-3089.2003.01140.x

Cited by 28 publications

(18 citation statements)

References 45 publications

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“…An approximately 750-bp fragment from the promoter region is reported to target variable levels of gene expression to the endothelium of capillaries, but not large vessels in transgenic mice. 27 This, together with the common use of distal regulatory elements to enhance tissue specific gene transcription, 28,29 points to the existence of regions outside the promoter fragment that enhance endogenous endoglin expression.…”

Section: Introductionmentioning

confidence: 99%

“…Prior work on the endoglin promoter has revealed that gene transcription is initiated from a stretch of DNA that has neither a TATA nor CAAT consensus sequence. 23,24 This promoter region contains GC-rich regions and consensus motifs for a number of transcription factors including Sp1, Ets, GATA, AP2, nuclear factor B (NFB), and TGF-␤, 27 This, together with the common use of distal regulatory elements to enhance tissue specific gene transcription, 28,29 points to the existence of regions outside the promoter fragment that enhance endogenous endoglin expression.In this study, we used comparative genomics to identify potential regulatory regions of endoglin. We show that conserved Ets binding sites are required for transcriptional activity of the promoter and identify Fli-1, Erg, and Elf-1 as 3 Ets family members that are bound to the promoter in endothelial cells in vivo.…”

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confidence: 99%

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Endoglin expression in the endothelium is regulated by Fli-1, Erg, and Elf-1 acting on the promoter and a –8-kb enhancer

Pimanda

Chan

Donaldson

et al. 2006

Blood

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Angiogenesis is critical to the growth and regeneration of tissue but is also a key component of tumor growth and chronic inflammatory disorders. Endoglin plays a key role in angiogenesis by modulating cellular responses to transforming growth factor-␤ (TGF-␤) signaling and is upregulated in proliferating endothelial cells. To gain insights into the transcriptional hierarchies that govern endoglin expression, we used a combination of comparative genomic, biochemical, and transgenic approaches. Both the promoter and a region 8 kb upstream of exon 1 were active in transfection assays in endothelial cells. In transgenic mice, the promoter directed low-level expression to a subset of endothelial cells. By contrast, inclusion of the ؊8 enhancer resulted in robust endothelial activity with additional staining in developing ear mesenchyme. Subsequent molecular analysis demonstrated that both the ؊8 enhancer and the promoter depend on conserved Ets sites, which were bound in endothelial cells in vivo by Fli-1, Erg, and Elf-1. This study therefore establishes the transcriptional framework within which endoglin functions during angiogenesis. IntroductionEndoglin is a transmembrane glycoprotein that is expressed predominantly in endothelial cells and is critical for the normal development of blood vessels. 1 Endoglin-null mice die in utero with defects in vascular remodeling, [2][3][4] and haploinsufficiency of the gene in humans is associated with type 1 hereditary hemorrhagic telangiectasia (HHT), an autosomal dominant disorder characterized by bleeding from vascular malformations in the skin, mucosa, and viscera (reviewed in Lebrin et al 5 and Abdalla and Letarte 6 ).Endoglin functions as an accessory receptor for members of the transforming growth factor-␤ (TGF-␤) superfamily of cytokines. 7 The TGF-␤ superfamily includes the TGF-␤ isoforms, nodal, activins, bone morphogenetic proteins (BMPs), and other related factors, which serve diverse functions during embryonic development and tissue homeostasis in the adult (reviewed in Massague 8 ). These cytokines assemble a primary receptor complex made up of type I and type II receptors on the surface of cells and control gene expression by receptor mediated phosphorylation of SMAD proteins. The type I receptors confer signal specificity by preferential phosphorylation of SMAD2/3 or SMAD1/5/8 isoforms, which broadly engage different binding partners and act on separate sets of target genes. Ancillary nonsignaling receptors such as endoglin regulate the pathway by selectively facilitating the binding of subsets of the TGF-␤ family members to the primary receptor complex. 9 Endoglin can also influence the cellular response to a particular cytokine by its preferential interaction with certain type I receptors. ALK1 is one such receptor, and mutations in this gene result in type 2 HHT, a clinical disorder that closely resembles that caused by mutations in the endoglin gene. 10 The endothelium is intimately involved in the development and progression of a number of pathologies, i...

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Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

Endoglin expression in the endothelium is regulated by Fli-1, Erg, and Elf-1 acting on the promoter and a –8-kb enhancer

Pimanda

Chan

Donaldson

et al. 2006

Blood

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“…However, the same promoter was unpredictable when driving the human A20 gene [19]. A study demonstrated that human intercellular adhesion molecule 2 (ICAM-2) promoter was much weaker in pig than in mice, suggesting that there may be a need for additional regulatory elements to achieve species-specific gene expression in pig [20]. Mouse major histocompatibility complex class I gene H-2K b promoter was used to express human TRAIL (tumor necrosis factor-alpharelated apoptosis-inducing ligand) in transgenic pigs.…”

Section: Discussionmentioning

confidence: 99%

Identification and cloning of the porcine ROSA26 promoter and its role in transgenesis

Li¹,

Burlak²,

Estrada³

et al. 2014

Transplant Technol

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Several promoters from different origins have been used to drive transgene expression in pigs with variable results. In this study, we identified porcine ROSA26 promoter located on chromosome 13. Porcine ROSA26 promoter was cloned and its activity was examined using enhanced green fluorescence protein (EGFP) as a reporter. EGFP expression was detected in adult liver derived cells (LDC) by flow cytometry and confocal microscopy after transient transfection. Stable LDC expressing EGFP were established and used as nuclear donors for somatic cell nuclear transfer (SCNT). Two hundred thirty-three reconstructed embryos were transferred to two recipients. One pregnancy was established. A total of four fetuses were collected at day 30 of gestation, one normal and three resorbed. All fetuses had EGFP gene integration. Confocal microscope analysis showed that EGFP was expressed in fetal liver, heart, and isolated fetal fibroblasts. These results indicated that ROSA26 promoter is a ubiquitous promoter and can be utilized in pig transgenesis.

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“…Various promoter sequences (ESS) were defined, responsible for endothelial-specific expression of these genes. However, as in the case of shear stress regulation, often more than one element was defined as an ESS in a single endogenous promoter, and the contribution of each of these sequences to endothelial specificity in vitro or in vivo (in the embryo or adult animals) has not been determined [32][33][34][35][36][37].…”

Section: Introductionmentioning

confidence: 99%

Shear stress-induced transcriptional regulation via hybrid promoters as a potential tool for promoting angiogenesis

et al. 2009

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Among the key effects of fluid shear stress on vascular endothelial cells is modulation of gene expression. Promoter sequences termed shear stress response elements (SSREs) mediate the responsiveness of endothelial genes to shear stress. While previous studies showed that shear stress responsiveness is mediated by a single SSRE, these endogenous promoters often encode for multiple SSREs. Moreover, hybrid promoters encoding a single SSRE rarely respond to shear stress at the same magnitude as the endogenous promoter. Thus, to better understand the interplay between the various SSREs, and between SSREs and endothelial-specific sequences (ESS), we generated a series of constructs regulated by SSREs cassettes alone, or in combination with ESS, and tested their response to shear stress and endothelial specific expression. Among these constructs, the most responsive promoter (NR1/2) encoded a combination of two GAGACC/SSREs, the Sp1/Egr1 sequence, as well as a TPA response element (TRE). This construct was four- to five-fold more responsive to shear stress than a promoter encoding a single SSRE. The expression of constructs containing other SSRE combinations was unaffected or suppressed by shear stress. Addition of ESS derived from the Tie2 promoter, either 5' or 3' to NR1/2 resulted in shear stress transcriptional suppression, yet retained endothelial specific expression. Thus, the combination and localization order of the various SSREs in a single promoter is crucial in determining the pattern and degree of shear stress responsiveness. These shear stress responsive cassettes may prove beneficial in our attempt to time the expression of an endothelial transgene in the vasculature.

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Targeting gene expression to endothelium in transgenic animals: a comparison of the human ICAM‐2, PECAM‐1 and endoglin promoters

Cited by 28 publications

References 45 publications

Endoglin expression in the endothelium is regulated by Fli-1, Erg, and Elf-1 acting on the promoter and a –8-kb enhancer

Endoglin expression in the endothelium is regulated by Fli-1, Erg, and Elf-1 acting on the promoter and a –8-kb enhancer

Identification and cloning of the porcine ROSA26 promoter and its role in transgenesis

Shear stress-induced transcriptional regulation via hybrid promoters as a potential tool for promoting angiogenesis

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