About Enhancer
| Enhancer ID: | E_01_263 | |
| Enhancer symbol: | -- | |
| Species: | Human | |
| Position : | chrX:55041578-55041679   |
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| Biosample name: | K-562 | |
| Experiment class : | Low throughput |
| Enhancer type: | Enhancer | |
| Disease: | -- | |
| DO: | -- | |
| Mesh: | -- | |
| Distance from TSS: | Intron | |
| Pubmed ID: | 28123038 |
| Enhancer experiment: | 3C,Luciferase Reporter Assay,RT-qPCR,CRISPR/Cas9 | |
| Enhancer experiment description: | In addition to this int-1-GATA site,another GATA site in ALAS2 intron 8 (int-8-GATA) has also been reported to function as an enhancer through luciferase assays (35). Furthermore, the physical proximity between the int-1-GATA and int-8-GATA sites and between the promoter and the int-8-GATA site were veriied by 3C assays (Figure 4H and I), suggesting that the promoter, the int-1-GATA site and the int-8-GATA site could form a potential chromatin enhancer loop. To further interrogate the speciic role of proximal int-1-GATA and distal int-8-GATA enhancer elements in ALAS2 regulation, we employed CRISPR/Cas9 technol-ogy to separately delete the GATA1 binding sites from the intron 1 or 8 enhancer regions in K562 cells.As expected,GATA site deletion in int1 6 and int8 4 cells entirely disrupted GATA1 occupancy at the intron 1 and 8 enhancer regions, respectively, whereas each deletion did not affect GATA1 occupancy at the other intronic GATA site and the promoter region (Figure 7C). We detected signiicant reductions in ALAS2 expression in both int1 6 and int8 4 cells during cytosine arabinoside (AraC)-inducedK562 erythroid differentiation (Fig-Nucleic Acids Research, 2017, Vol. 45, No. 2 667 ure 7D and E). |
About Target gene
| Target gene : | ALAS2(ALAS-E,ALASE,ANH1,ASB,SIDBA1,XLDPP,XLEPP,XLSA) | |
| Strong evidence: | 3C,CRISPR/Cas9 | |
| Less strong evidence: | Luciferase Reporter Assay,RT-qPCR | |
| Target gene experiment description: | In addition to this int-1-GATA site,another GATA site in ALAS2 intron 8 (int-8-GATA) has also been reported to function as an enhancer through luciferase assays (35). Furthermore, the physical proximity between the int-1-GATA and int-8-GATA sites and between the promoter and the int-8-GATA site were veriied by 3C assays (Figure 4H and I), suggesting that the promoter, the int-1-GATA site and the int-8-GATA site could form a potential chromatin enhancer loop. To further interrogate the speciic role of proximal int-1-GATA and distal int-8-GATA enhancer elements in ALAS2 regulation, we employed CRISPR/Cas9 technol-ogy to separately delete the GATA1 binding sites from the intron 1 or 8 enhancer regions in K562 cells.As expected,GATA site deletion in int1 6 and int8 4 cells entirely disrupted GATA1 occupancy at the intron 1 and 8 enhancer regions, respectively, whereas each deletion did not affect GATA1 occupancy at the other intronic GATA site and the promoter region (Figure 7C). We detected signiicant reductions in ALAS2 expression in both int1 6 and int8 4 cells during cytosine arabinoside (AraC)-inducedK562 erythroid differentiation (Fig-Nucleic Acids Research, 2017, Vol. 45, No. 2 667 ure 7D and E). |
About TF
| TF name : | GATA1 (ERYF1,GATA-1,GF-1,GF1,NF-E1,NFE1,XLANP,XLTDA,XLTT) | |
| TF experiment: | ChIP-qPCR | |
| TF experiment description: | The ChIP-qPCR assay showed the binding of GATA1 to the promoter and the intron 1 and 8 enhancer regions in int1_x0002_6, int8_x0002_4 and WT K562 cells. The DNA sequence at the exon 4-intron 4 junction (exon 4/intron 4) acts as a negative control region. Normal rabbit IgG was employed as the control in all ChIP assays. |
About Function
| Enhancer function : | -- |
| Enhancer function experiment: | -- |
| Enhancer function experiment description: |
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About SNP
| SNP ID: | -- | |
| SNP position: | -- |
| SNP experiment: | -- |
Enhancer associated network
The number on yellow line represents the distance between enhancer and target gene
