About Enhancer
| Enhancer ID: | E_01_274 | |
| Enhancer symbol: | -- | |
| Species: | Human | |
| Position : | chr1:221049243-221051243   |
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| Biosample name: | HUVEC | |
| Experiment class : | Low+High throughput |
| Enhancer type: | Enhancer | |
| Disease: | -- | |
| DO: | -- | |
| Mesh: | -- | |
| Distance from TSS: | >2KB | |
| Pubmed ID: | 28536097 |
| Enhancer experiment: | ChIP-seq,Luciferase Reporter Assay,ChIP | |
| Enhancer experiment description: | We also performed H3K27ac ChIP-seq and found that 94% of conserved ERG peaks overlapped H3K27ac-enriched regions, supporting their association with active enhancers. We also identified an ERG-bound enhancer, conserved in cows and humans, ~3.0 kb upstream of the gene H2.0-Like Homeobox (HLX) (Fig. 8B; Fig. S8). We cloned this conserved H3K27ac- and ERG-enriched -3 kb 5’ putative regulatory region (HLX-3a, 1565 bp fragment) upstream of a minimal promoter (SV40) driving a luciferase reporter, and found that it was VEGF-responsive, and that the basal and VEGF-induced activity of this enhancer required ETS DNA binding sequences (Fig. 9D). |
About Target gene
| Target gene : | HLX(HB24,HLX1) | |
| Strong evidence: | -- | |
| Less strong evidence: | ChIP-seq,Luciferase Reporter Assay | |
| Target gene experiment description: | We also performed H3K27ac ChIP--seq and found that 94% of conserved ERG peaks overlapped H3K27ac--enriched regions, supporting their association with active enhancers. We also identified an ERG--bound enhancer, conserved in cows and humans, ~3.0 kb upstream of the gene H2.0--Like Homeobox (HLX) (Fig. 8B; Fig. S8). We cloned this conserved H3K27ac-- and ERG--enriched --3 kb 5’ putative regulatory region (HLX--3a, 1565 bp fragment) upstream of a minimal promoter (SV40) driving a luciferase reporter, and found that it was VEGF--responsive, and that the basal and VEGF--induced activity of this enhancer required ETS DNA binding sequences (Fig. 9D). |
About TF
| TF name : | VEGF(VEGF)ERG(erg-3,p55) | |
| TF experiment: | Luciferase Reporter Assay | |
| TF experiment description: | We cloned this conserved H3K27ac- and ERG-enriched -3 kb 5’ putative regulatory region (HLX-3a, 1565 bp fragment) upstream of a minimal promoter (SV40) driving a luciferase reporter, and found that it was VEGF-responsive, and that the basal and VEGF-induced activity of this enhancer required ETS DNA binding sequences (Fig. 9D). |
About Function
| Enhancer function : | A highly-conserved ERG-bound regulatory element was required in the VEGF responsiveness of the angiogenic gene, HLX. |
| Enhancer function experiment: | CRISPR/Cas9,PCR |
| Enhancer function experiment description: |
To further test the functional importance of this enhancer, we utilized CRISPR genome editing to delete a portion (1201 bp; see Fig. 8B for schematic) of the H3K27Ac-enriched, ERG-bound region upstream of HLX in TeloHAECs, an immortalized aortic EC line. Several clonal lines (ΔHLX15, ΔHLX17 and ΔHLX21) heterozygous for deletion of this region were generated and confirmed by PCR and DNA sequencing (data not shown). Comparison was made to a clonal line generated following transfection of scrambled control gRNAs (Scr3). While the basal expression of HLX appeared to be unaffected in the deletion lines, the VEGF-dependent induction of HLX was attenuated (Fig. 9G). In contrast, DLL4 induction was unaffected. Furthermore, knock-down of ERG appeared to attenuate the induction of HLX to a greater extent in the control line compared to the deletion lines, implying that ERG acts through the deleted enhancer region. Collectively, these findings demonstrate the requirement of a highly-conserved ERG-bound regulatory element in the VEGF responsiveness of the angiogenic gene, HLX. |
About SNP
| SNP ID: | -- | |
| SNP position: | -- |
| SNP experiment: | -- |
Enhancer associated network
The number on yellow line represents the distance between enhancer and target gene
