About Enhancer
| Enhancer ID: | E_01_273 | |
| Enhancer symbol: | -- | |
| Species: | Human | |
| Position : | chr11:355447-358949   |
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| Biosample name: | A549,HEK-293 | |
| Experiment class : | Low throughput |
| Enhancer type: | Enhancer | |
| Disease: | -- | |
| DO: | -- | |
| Mesh: | -- | |
| Distance from TSS: | >2KB | |
| Pubmed ID: | 28511927 |
| Enhancer experiment: | Luciferase Reporter Assay,CRISPR/Cas9,ChIP,EMSA,3C | |
| Enhancer experiment description: | Here, through public data mining, episomal luciferase reporter assay and in vivo CRISPR-Cas9 genome editing, we identified an IFN-responsive enhancer located 35 kb upstream of IFITM3 gene promoter upregulating the IFN-induced expression of IFITM1, 2 and 3 genes. Chromatin immunoprecipitation (ChIP), electrophoretic mobility shift assay (EMSA) and luciferase reporter assay demonstrated that signal transducers and activators of transcription (STAT) 1 bound to the enhancer with the treatment of IFN and was indispensable for the enhancer activity. Furthermore, using chromosome conformation capture technique, we revealed that the IFITM1, 2 and 3 genes physically clustered together and constitutively looped to the distal enhancer through long-range interactions in both HEK293 and A549 cells, providing structural basis for coordinated regulation of IFITM1, 2 and 3 by the enhancer. Finally, we showed that in vivo truncation of the enhancer impaired IFN-induced resistance to influenza A virus (IAV) infection. |
About Target gene
| Target gene : | IFITM1(9-27,CD225,DSPA2a,IFI17,LEU13),IFITM2(1-8D,DSPA2c),IFITM3(1-8U,DSPA2b,IP15) | |
| Strong evidence: | CRISPR/Cas9,3C | |
| Less strong evidence: | Luciferase Reporter Assay,EMSA | |
| Target gene experiment description: | Here, through public data mining, episomal luciferase reporter assay and in vivo CRISPR-Cas9 genome editing, we identified an IFN-responsive enhancer located 35 kb upstream of IFITM3 gene promoter upregulating the IFN-induced expression of IFITM1, 2 and 3 genes. Chromatin immunoprecipitation (ChIP), electrophoretic mobility shift assay (EMSA) and luciferase reporter assay demonstrated that signal transducers and activators of transcription (STAT) 1 bound to the enhancer with the treatment of IFN and was indispensable for the enhancer activity. Furthermore, using chromosome conformation capture technique, we revealed that the IFITM1, 2 and 3 genes physically clustered together and constitutively looped to the distal enhancer through long-range interactions in both HEK293 and A549 cells, providing structural basis for coordinated regulation of IFITM1, 2 and 3 by the enhancer. |
About TF
| TF name : | STAT1(CANDF7,IMD31A,IMD31B,IMD31C,ISGF-3,STAT91) | |
| TF experiment: | ChIP,EMSA,3C | |
| TF experiment description: | Chromatin immunoprecipitation (ChIP), electrophoretic mobility shift assay (EMSA) and luciferase reporter assay demonstrated that signal transducers and activators of transcription (STAT) 1 bound to the enhancer with the treatment of IFN and was indispensable for the enhancer activity. Furthermore, using chromosome conformation capture technique, we revealed that the IFITM1, 2 and 3 genes physically clustered together and constitutively looped to the distal enhancer through long-range interactions in both HEK293 and A549 cells, providing structural basis for coordinated regulation of IFITM1, 2 and 3 by the enhancer. |
About Function
| Enhancer function : | In vivo truncation of the enhancer impaired IFN-induced resistance to influenza A virus (IAV) infection. |
| Enhancer function experiment: | RT-PCR,Western blot |
| Enhancer function experiment description: |
As IFITM1, 2 and 3 are very important for host to resist infection of IAV [4], [8], we infected wild-type and E2-3-truncated HEK293 cells with IAV Puerto Rico/8/1934 (PR8) at a MOI of 0.1 for 24 h. Although IFITM1, 2 and 3 were induced by PR8 virus in both wild-type and E2-3-truncated HEK293 cells, the inductions were greatly impaired upon truncation of E2-3 (Fig. 5A and B), further confirming the regulation of IFITM1, 2 and 3 expression by the enhancer E2-3. Surprisingly, we did not observe a significant effect on the replication of IAV when enhancer E2-3 was truncated, as revealed by the mRNA and protein levels of virus NP and progeny virus titers determined by TCID50 assay (Fig. S9). Nonetheless, when we treated wild-type and E2-3-truncated HEK293 cells with IFNβ respectively for 24 h before infection of PR8, NP mRNA and protein levels were found to be much higher in mutant HEK293 cells (Fig. 5C and D). Moreover, consistent with virus NP expression, the viral titers were also increased in E2-3-truncated cells (Fig. 5E), indicating that enhancer E2-3 was responsible for IFNβ-induced resistance to IAV infection. |
About SNP
| SNP ID: | -- | |
| SNP position: | -- |
| SNP experiment: | -- |
Enhancer associated network
The number on yellow line represents the distance between enhancer and target gene
