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| Status |
Public on Nov 10, 2020 |
| Title |
Genome-wide investigation of the dynamic changes of epigenome modifications after DNA methylation editing [ChIP-Seq] |
| Organism |
Homo sapiens |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
The aim of this study was to investigate the stability of DNA methylation, the dynamics of different histone modifications and the changes in expression after genome-wide introduction of DNA methylation in promotor CpG islands. Therefore, the catalytic domain of DNMT3A was fused to a zinc finger protein and expressed for three days to globally introduce de novo DNA methylation and subsequently, the changes in the epigenome were monitored for up to eleven days.
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| Overall design |
A stable HEK293 cell line was constructed with the catalytic domain of DNMT3A fused to a zinc finger (ZnF-3AC) which is under the control of a doxycycline (dox)-inducible promoter. GFP is co-expressed via an IRES and serves as a reporter. Cells were either harvested before dox treatment ("no dox") or grown under dox conditions for three days ("3d dox"). GFP-positive cells were enriched by FACS after 3d dox and a fraction of these cells was re-seeded on a 6-Well-plate without supplementation of dox. Five and eight days after dox removal ("5d / 8d / 11d off"), cells were harvested and a fraction of cells was re-seeded again. After eleven days of dox removal ("11d off"), cells were harvested and the experiment was stopped.
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| Contributor(s) |
Broche J, Kungulovski G, Bashtrykov P, Rathert P, Jeltsch A |
| Citation(s) |
33300025 |
| |
| Submission date |
Feb 07, 2020 |
| Last update date |
Jan 19, 2021 |
| Contact name |
Albert Jeltsch |
| E-mail(s) |
[email protected]
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| Phone |
+49 685 64390
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| Organization name |
University Stuttgart
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| Department |
Institute of Biochemistry and Technical Biochemistry
|
| Street address |
Allmandring 31
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| City |
Stuttgart |
| ZIP/Postal code |
70569 |
| Country |
Germany |
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| Platforms (2) |
| GPL16791 |
Illumina HiSeq 2500 (Homo sapiens) |
| GPL21290 |
Illumina HiSeq 3000 (Homo sapiens) |
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| Samples (24)
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| This SubSeries is part of SuperSeries: |
| GSE144904 |
Genome-wide investigation of the dynamic changes of epigenome modifications after DNA methylation editing |
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| Relations |
| BioProject |
PRJNA605353 |
| SRA |
SRP247630 |
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