About Enhancer
| Enhancer ID: | E_01_005 | |
| Enhancer symbol: | p53enh3507 | |
| Species: | Human | |
| Position : | chr6:36631690-36640853   |
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| Biosample name: | BJ | |
| Experiment class : | Low+High throughput |
| Enhancer type: | Enhancer | |
| Disease: | -- | |
| DO: | -- | |
| Mesh: | -- | |
| Distance from TSS: | >2KB | |
| Pubmed ID: | 26751173 |
| Enhancer experiment: | ChIP-seq,CRISPR/Cas9,Luciferase Reporter Assay,siRNA,GRO-seq | |
| Enhancer experiment description: | We identified several functional enhancer elements and characterized the role of two of them in mediating p53 (TP53) and ERa (ESR1) gene regulation. To build a CRISPR-Cas9 single guide RNA (sgRNA) library, we followed a strategy that enabled us to target ≈90% of p53-bound enhancers ( (Fig. 1b and Supplementary Notably, two independent sgRNAs targeted a putative enhancer located ~10 kb upstream of CDKN1A (formerly known as p21), which is a key effector of p53-dependent OIS (Fig. 1e; p53enh3507). Another top-scoring sgRNA mapped to a known p53-responsive element that is located proximal to the transcription start site of CDKN1A (Fig. 1e; p53enh3508). Next, we assessed the enhancer capacity of the p53enh3507 region by cloning it into a reporter vector and verified that it strongly induces transcription (Fig. 2a). We also observed a substantial reduction in enhancing activity upon siRNA-mediated knockdown of p53 (Fig. 2b). We obtained similar results for the p53enh3508 enhancer element (Supplementary Fig. 2b,c). |
About Target gene
| Target gene : | CDKN1A(CAP20,CDKN1,CIP1,MDA-6,P21,SDI1,WAF1,p21CIP1) | |
| Strong evidence: | -- | |
| Less strong evidence: | Luciferase Reporter Assay | |
| Target gene experiment description: | Next, we assessed the enhancer capacity of the p53enh3507 region by cloning it into a reporter vector and verified that it strongly induces transcription (Fig. 2a). We also observed a substantial reduction in enhancing activity upon siRNA-mediated knockdown of p53 (Fig. 2b). We obtained similar results for the p53enh3508 enhancer element (Supplementary Fig. 2b,c). Interestingly,concurrent disruption of TP53enh3507 and TP53enh3508 resulted in further reduction of CDKN1A expression,suggesting an independent regulation by both Enhancer elements. |
About TF
| TF name : | TP53(BCC7,BMFS5,LFS1,P53,TRP53) | |
| TF experiment: | Luciferase Reporter Assay | |
| TF experiment description: | The same assay as in a, only that cells were co-transfected with control, or p53-targeting short interfering RNA (si-Cont.; si-p53). A reporter vector containing the enhancer region p53-BER4 was used as a positive control for p53-dependency. The efficiency of p53 knockdown was determined by immunoblot analysis. |
About Function
| Enhancer function : | The p53enh3507 is an endogenous Enhancer of CDKN1A and disruption of this region causes bypass of senescence in p53-WT cells. |
| Enhancer function experiment: | BrdU Assay,β-galactosidase Assay |
| Enhancer function experiment description: |
Upon oncogene activation, one major function of p53 is to activate an irreversible cell-cycle arrest program named oncogene-induced senescence (OIS). We performed BrdU labeling and senescence-associated β-galactosidase (β-gal) assays and found that sgRNA-p53enh3507 and sgRNA-p53enh3508 caused OIS bypass and continuous cell proliferation (Fig. 1g,h and Supplementary Fig. 2a) Collectively, these results demonstrate that p53enh3507 is an endogenous Enhancer of CDKN1A and disruption of this region causes bypass of senescence in p53-WT cells.Moreover, cooperative action of p53enh3507 (distal Enhancer of CDKN1A-deCD-KN1A) and p53enh3508 (proximal Enhancer of CDKN1A-peCDKN1A) is required to activate CDKN1A expression and initiate OIS. |
About SNP
| SNP ID: | -- | |
| SNP position: | -- |
| SNP experiment: | -- |
Enhancer associated network
The number on yellow line represents the distance between enhancer and target gene
