Enhancer ID: | E_01_354 | |
Enhancer symbol: | POU5F1 distal Enhancer | |
Species: | Human | |
Position : | chr6:31140556-31141204 |
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Biosample name: | Human Primordial Germ Cell | |
Experiment class : | Low+High throughput |
Enhancer type: | Enhancer | |
Disease: | -- | |
DO: | -- | |
Mesh: | -- | |
Distance from TSS: | >2KB | |
Pubmed ID: | 30590035 |
Enhancer experiment: | ATAC-seq,ChIP-seq,ChIP-qPCR | |
Enhancer experiment description: | Using chromatin immunoprecipitation sequencing (ChIP-seq) for TFAP2C in 5i/L/FA cultured naive hPSCs (Pastor et al., 2018), we found that the 1,892 hPGC and hPGCLC overlapping peaks are also bound by TFAP2C in 5i/L/FA cultured cells (Figure 3C).Analysis of the 1,892 peaks enriched in the naive-specific and germline cell-specific intersect group revealed a significant enrichment of AP2 motifs, as expected (Figure S3B).This observation raises the possibility that restriction of OCT4 expression between days 2 and 3 of aggregate differentiation may be due to NE and/or the DE enhancer activation at the OCT4 locus. Given that the NE is bound by TFAP2C in ground-state naive pluripotent stem cells, whereas the DE is not (Figure 5C), we next confirmed that the OCT4 NE is also a target of TFAP2C during hPGCLC differentiation. To do this, we performed ChIP-qPCR on day 4 aggregates containing hPGCLCs and discovered that TFAP2C is bound to the NE, whereas it is not bound to a genomic region that does not contain AP2 sites (Figure 5D). Taken together, these experiments suggest that OCT4 regulation during hPGCLC differentiation may involve enhancer activation at the DE as well as TFAP2C-bound enhancer activation at the NE. |
Target gene : | POU5F1(OCT3,OCT4,OTF-3,OTF3,OTF4,Oct-3,Oct-4) | |
Strong evidence: | -- | |
Less strong evidence: | ChIP-qPCR | |
Target gene experiment description: | Screenshot of ATAC-seq and TFAP2C ChIP-seq signals showing three Enhancers at the POU5F1 locus (encoding OCT4). Shaded boxes highlight the naive Enhancer(NE),proximalEnhancer(PE),anddistalEnhancer(DE)atthePOU5F1locus.DEdeletion andNEdeletion indicategenomicregionsthatweredeleted by CRISPR/Cas9-mediated genome editing. Primers for ChIP-qPCR (P1-2,P3-4,P5-6) of NE and control regions are shown. |
TF name : | TFAP2C(AP2-GAMMA,ERF1,TFAP2G,hAP-2g) | |
TF experiment: | CRISPR/Cas9 | |
TF experiment description: | Using CRISPR/Cas9, we show that deleting the TFAP2C-bound naive Enhancer at the OCT4 locus (also called POU5F1) results in impaired OCT4 expression and a negative effect on hPGCLC identity. |
Enhancer function : | OCT4 regulation during hPGCLC differentiation may involve enhancer activation at the DE as well as TFAP2C-bound enhancer activation at the NE. |
Enhancer function experiment: | ATAC-seq,ChIP-qPCR |
Enhancer function experiment description: |
Taken together, these experiments suggest that OCT4 regulation during hPGCLC differentiation may involve enhancer activation at the DE as well as TFAP2C-bound enhancer activation at the NE. |
SNP ID: | -- | |
SNP position: | -- |
SNP experiment: | -- |
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