|
| Status |
Public on Apr 22, 2020 |
| Title |
Looping of upstream cis-regulatory elements is required for CFTR expression in human airway epithelial cells [ChIP-seq] |
| Organism |
Homo sapiens |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
The CFTR gene lies within an invariant topologically associated domain (TAD) demarcated by CTCF and cohesin, but shows cell-type specific control mechanisms utilizing different cis-regulatory elements (CRE) within the TAD. Within the respiratory epithelium, more than one cell type expresses CFTR and the molecular mechanism controlling its transcription are likely divergent between them. Here we determine how two extragenic CREs that are prominent in secretory epithelial cells in the lung, regulate expression of the gene. We showed earlier that these CREs, located at -44kb and -35 kb upstream of the promoter, have strong cell-type-selective enhancer function. They are also responsive to inflammatory mediators and to oxidative stress, consistent with a key role in CF lung disease. Here we use CRISPR/Cas9 technology to remove these CREs from the endogenous locus in human bronchial epithelial cells. Loss of either site extinguished CFTR expression and abolished long-range interactions between these sites and the gene promoter. The deletions also greatly reduced promoter interactions with the 5’ TAD boundary. We show substantial recruitment of RNAPII to the -35kb element and identify CEBPβ as a key activating transcription factor for airway expression of CFTR, likely through occupancy at both the CRE and the gene promoter.
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| |
|
| Overall design |
Examination of H3K27Ac and RNA Polymerase II occupancy in 16HBE14o-, Caco2 and Calu3 cells
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| |
|
| Contributor(s) |
Harris A, NandyMazumdar M, Paranjapye A |
| Citation(s) |
32095812 |
| |
| Submission date |
Jun 16, 2019 |
| Last update date |
Apr 22, 2020 |
| Contact name |
Ann Harris |
| E-mail(s) |
axh575@case.edu
|
| Organization name |
Case Western Reserve Uni
|
| Street address |
10900 Euclid Ave
|
| City |
Cleveland |
| State/province |
OH |
| ZIP/Postal code |
44106 |
| Country |
USA |
| |
|
| Platforms (2) |
| GPL16791 |
Illumina HiSeq 2500 (Homo sapiens) |
| GPL20301 |
Illumina HiSeq 4000 (Homo sapiens) |
|
| Samples (10)
|
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| This SubSeries is part of SuperSeries: |
| GSE132808 |
Looping of upstream cis-regulatory elements is required for CFTR expression in human airway epithelial cells |
|
| Relations |
| BioProject |
PRJNA549087 |
| SRA |
SRP201553 |
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