Reference - Detail
| RRC ID | 63661 |
|---|---|
| Author | Tsujimura T, Takase O, Yoshikawa M, Sano E, Hayashi M, Hoshi K, Takato T, Toyoda A, Okano H, Hishikawa K. |
| Title | Controlling gene activation by enhancers through a drug-inducible topological insulator. |
| Journal | Elife |
| Abstract |
While regulation of gene-enhancer interaction is intensively studied, its application remains limited. Here, we reconstituted arrays of CTCF-binding sites and devised a synthetic topological insulator with tetO for chromatin-engineering (STITCH). By coupling STITCH with tetR linked to the KRAB domain to induce heterochromatin and disable the insulation, we developed a drug-inducible system to control gene activation by enhancers. In human induced pluripotent stem cells, STITCH inserted between MYC and the enhancer down-regulated MYC. Progressive mutagenesis of STITCH led to a preferential escalation of the gene-enhancer interaction, corroborating the strong insulation ability of STITCH. STITCH also altered epigenetic states around MYC. Time-course analysis by drug induction uncovered deposition and removal of H3K27me3 repressive marks follows and reflects, but does not precede and determine, the expression change. Finally, STITCH inserted near NEUROG2 impaired the gene activation in differentiating neural progenitor cells. Thus, STITCH should be broadly useful for functional genetic studies. |
| Volume | 9 |
| Published | 2020-5-5 |
| DOI | 10.7554/eLife.47980 |
| PII | 47980 |
| PMID | 32369019 |
| PMC | PMC7200164 |
| MeSH | CCCTC-Binding Factor / drug effects CCCTC-Binding Factor / genetics CCCTC-Binding Factor / metabolism Chromatin / drug effects* Chromatin / metabolism Enhancer Elements, Genetic / genetics* Epigenesis, Genetic / drug effects Epigenesis, Genetic / genetics Gene Expression Regulation / drug effects* Genes / genetics* Genetic Engineering / methods Humans Induced Pluripotent Stem Cells / drug effects Induced Pluripotent Stem Cells / metabolism Proto-Oncogene Proteins c-myc / genetics Proto-Oncogene Proteins c-myc / metabolism Transcriptional Activation / drug effects Transcriptional Activation / genetics Transcriptome |
| IF | 7.08 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 14 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | 253G1(HPS0002) |