| Autor: |
Yang Liu, Xuetong Yue, Haizhu Zhang, Zhaoyu Qin, Fuchu He, Li Kai, Jiange Qiu, Wenjun Yang, Xiaogang Sun, Ziyan Xu, Jinwen Feng, Chen Ding, Jiangyan Zhao |
| Rok vydání: |
2021 |
| Předmět: |
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| Zdroj: |
Science advances. 7(43) |
| ISSN: |
2375-2548 |
| Popis: |
To directly and quantitatively identify the transcriptional protein complexes assembled on accessible chromatin, we develop an assay for transposase-accessible chromatin using mass spectrum (ATAC-MS) based on direct transposition of biotinylated adaptors into open chromatin. Coupling with activated gene sequence information by ATAC-seq, ATAC-MS can profile the accessible chromatin-protein machinery. ATAC-MS, combined with fractionation strategies (fATAC-MS), can provide a high-resolution chromatin-transcriptional machinery atlas. ATAC-MS with a novel Tn5-dCas9 fusion protein [dCas9-targeted ATAC-MS (ctATAC-MS)] further facilitates systematic pinpointing of the transcriptional machinery at specific open chromatin regions. We used ATAC-MS and ATAC-seq to investigate transcriptional regulation during C2C12 cell differentiation and demonstrated the role of RFX1 in regulating the proliferation and differentiation of C2C12 cells. Our strategy provides a universal toolbox including ATAC-MS, fATAC-MS, and ctATAC-MS, which enables us to portray the transcriptional regulation machinery atlas in genome scale and investigate the protein-DNA complex at a specific genomic locus. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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