Inhibition of Interleukin-4- and CD40-induced IgE Germline Gene Promoter Activity by 2′-Aminoethoxy-modified Triplex-forming Oligonucleotides

Stütz, Adrian M.; Hoeck, Jutta; Natt, François; Cuenoud, Bernard; Woisetschläger, Maximilian

doi:10.1074/jbc.m010260200

Cited by 26 publications

(19 citation statements)

References 44 publications

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“…Here we demonstrate the ability of antiparallel purine motif TFOs to selectively form DNA triplex at two sequences encoding tandem Ets core DNA binding motifs in the tie-1 promoter and the triplex-mediated inhibition of promoter activity through TFO binding to one of these sequences (E-1). Although the inhibition of the IgE germline proximal gene promoter through triplex DNA formation at a sequence regulated by STAT6, NF-κB, and the Ets factor PU.1 was reported recently (30), to our knowledge this is the first demonstration of selective triplex DNA formation at sequences containing multiple Ets binding sites.…”

Section: Discussionmentioning

confidence: 99%

“…Triplex DNA formation has been investigated as a highly selective means of regulating many clinically relevant targets such as c-myc (25,31), cyclin D1 (24), mdr-1 (27), HIV (28), insulin-like growth factor-1 receptor (29), platelet-derived growth factor (32), rhodopsin (53), and the IgE germline gene promoter (30). In addition to the disruption of transcriptional elongation (21,22,(27)(28)(29)53), inhibition of gene expression has been achieved through triplex DNA formation at sequences encoding transcription factor binding sites including SP-1, SRF, CNBP, PuF, MAZ, Pax5, PU.1, STAT6, and NF-κB (21,22,(24)(25)(26)31,32).…”

Section: Discussionmentioning

confidence: 99%

“…TFO can be directed to inhibit gene expression by competing with the binding of activating transcription factors to regulatory sites or by disrupting transcriptional elongation (21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32). Advances in this approach have been achieved through the development of oligonucleotide analogs such as N3′ → P5′ phosphoramidates (28), peptide nucleic acid (PNA) (33), and 2′-aminoethoxy-substituted riboses (30) that are resistant to intra-and extracellular nucleases and form strong DNA triplexes (21)(22)(23). Whereas the close association of condensed DNA with chromatin can serve as a barrier to DNA triplex formation, this does not appear to present a problem when targeting transcriptionally active regions of DNA (21).…”

Section: Introductionmentioning

confidence: 99%

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Selective Inhibition of the Human tie-1 Promoter with Triplex-Forming Oligonucleotides Targeted to Ets Binding Sites

Hewett

Daft

Laughton

et al. 2006

Mol Med

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The Tie receptors (Tie-1 and Tie-2/Tek) are essential for angiogenesis and vascular remodeling/integrity. Tie receptors are upregulated in tumor-associated endothelium, and their inhibition disrupts angiogenesis and can prevent tumor growth as a consequence. To investigate the potential of anti-gene approaches to inhibit tie gene expression for anti-angiogenic therapy, we have examined triple-helical (triplex) DNA formation at 2 tandem Ets transcription factor binding motifs (designated E-1 and E-2) in the human tie-1 promoter. Various tie-1 promoter deletion/mutation luciferase reporter constructs were generated and transfected into endothelial cells to examine the relative activities of E-1 and E-2. The binding of antiparallel and parallel (control) purine motif oligonucleotides (21-22 bp) targeted to E-1 and E-2 was assessed by plasmid DNA fragment binding and electrophoretic mobility shift assays. Triplex-forming oligonucleotides were incubated with tie-1 reporter constructs and transfected into endothelial cells to determine their activity. The Ets binding motifs in the E-1 sequence were essential for human tie-1 promoter activity in endothelial cells, whereas the deletion of E-2 had no effect. Antiparallel purine motif oligonucleotides targeted at E-1 or E-2 selectively formed strong triplex DNA (K d~1 0 -7 M) at 37 °C. Transfection of tie-1 reporter constructs with triplex DNA at E-1, but not E-2, specifically inhibited tie-1 promoter activity by up to 75% compared with control oligonucleotides in endothelial cells. As similar multiple Ets binding sites are important for the regulation of several endothelial-restricted genes, this approach may have broad therapeutic potential for cancer and other pathologies involving endothelial proliferation/dysfunction.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Selective Inhibition of the Human tie-1 Promoter with Triplex-Forming Oligonucleotides Targeted to Ets Binding Sites

Hewett

Daft

Laughton

et al. 2006

Mol Med

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“…The labeled product was purified by size exclusion chromatography (G-25 Sephadex Microspin columns; Amersham Life Sciences) and polyacrylamide gel electrophoresis. The binding reaction with equal amounts of whole-cell extract was performed as described [45]. For supershift experiments, 1 lg of specific antibody was pre-incubated on ice for 30 min before addition of approximately 0.5 ng labeled ds-DNA and further incubation for 30 min on ice.…”

Section: Cells Cell Culture Antibodies Proliferationmentioning

confidence: 99%

CD45 isoform expression is associated with different susceptibilities of human naive and effector CD4⁺ T cells to respond to IL‐4

et al. 2005

Self Cite

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Interleukin-4 (IL-4) is the major factor promoting the development of T helper type 2 (Th2) cells from naive precursor T cells. Minute amounts of IL-4 produced by naive T cells seem to be sufficient; however, the molecular mechanisms explaining this efficient utilization of are not yet known. Here, we show that human CD4 + CD45RA + naive T cells, in contrast to CD4 + CD45R0 + effector T cells, show responsiveness to endogenous as well as exogenous IL-4 to proliferate and differentiate towards Th2 cells in vitro. Despite production levels of IL-4 below conventional detection limits, CD45RA + T cell-derived IL-4 could clearly activate STAT6. Although the expression levels of IL-4R and STAT6 were not different between naive and effector T cells, only naive T cells responded to IL-4 in a STAT6-dependent reporter gene assay. Transfecting a trans-dominant negative form of STAT6 abrogated IL-4-induced proliferation in CD45RA + cells. A significantly higher protein tyrosine phosphatase (PTPase) activity was detected in CD45R0 + T cells as compared to CD45RA + T cells. Cross-linking CD45 potently reduced PTPase activity in CD45R0 + T cells and restored their ability to proliferate in response to IL-4. Thus, CD45 PTPase activity contributes to the susceptibility of naive and memory T cells to respond to IL-4.

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“…A positive charge and an RNA-like sugar conformation have been joined in the 2′-O-(2-aminoethyl) (AE) ribose derivatives developed by Cuenoud and colleagues (55)(56)(57). TFOs carrying these substitutions show enhanced kinetics of triplex formation and greater stability of the resultant complex at physiological pH and low Mg ++ concentration.…”

Section: Oligonucleotide Modifications Improve Tfo Activity Under Phymentioning

confidence: 99%

The potential for gene repair via triple helix formation

Seidman¹,

Glazer²

2003

J. Clin. Invest.

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Triplex-forming oligonucleotides (TFOs) can bind to polypurine/polypyrimidine regions in DNA in a sequence-specific manner. The specificity of this binding raises the possibility of using triplex formation for directed genome modification, with the ultimate goal of repairing genetic defects in human cells. Several studies have demonstrated that treatment of mammalian cells with TFOs can provoke DNA repair and recombination, in a manner that can be exploited to introduce desired sequence changes. This review will summarize recent advances in this field while also highlighting major obstacles that remain to be overcome before the application of triplex technology to therapeutic gene repair can be achieved

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Inhibition of Interleukin-4- and CD40-induced IgE Germline Gene Promoter Activity by 2′-Aminoethoxy-modified Triplex-forming Oligonucleotides

Cited by 26 publications

References 44 publications

Selective Inhibition of the Human tie-1 Promoter with Triplex-Forming Oligonucleotides Targeted to Ets Binding Sites

Selective Inhibition of the Human tie-1 Promoter with Triplex-Forming Oligonucleotides Targeted to Ets Binding Sites

CD45 isoform expression is associated with different susceptibilities of human naive and effector CD4⁺ T cells to respond to IL‐4

The potential for gene repair via triple helix formation

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Inhibition of Interleukin-4- and CD40-induced IgE Germline Gene Promoter Activity by 2′-Aminoethoxy-modified Triplex-forming Oligonucleotides

Cited by 26 publications

References 44 publications

Selective Inhibition of the Human tie-1 Promoter with Triplex-Forming Oligonucleotides Targeted to Ets Binding Sites

Selective Inhibition of the Human tie-1 Promoter with Triplex-Forming Oligonucleotides Targeted to Ets Binding Sites

CD45 isoform expression is associated with different susceptibilities of human naive and effector CD4+ T cells to respond to IL‐4

The potential for gene repair via triple helix formation

Contact Info

Product

Resources

About

CD45 isoform expression is associated with different susceptibilities of human naive and effector CD4⁺ T cells to respond to IL‐4