Protocol for visualizing the chromatin assembly properties of epigenetic protein complexes via an HTM module-mediated artificial tethering system
Summary: The detailed chromatin assembly processes for many epigenetic regulatory complexes are largely unknown. Here, we present a protocol utilizing heterochromatin-targeting module (HTM) module-mediated chromatin tethering followed by microscopy-based visualization to detect the recruitment prior...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725000036 |
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| Summary: | Summary: The detailed chromatin assembly processes for many epigenetic regulatory complexes are largely unknown. Here, we present a protocol utilizing heterochromatin-targeting module (HTM) module-mediated chromatin tethering followed by microscopy-based visualization to detect the recruitment priority between two components in Polycomb repressive complex 1 (PRC1). Moreover, we detail procedures for detecting the resultant histone-modifying activities of PRC1 using immunofluorescence (IF) analyses. This approach allows directly visualization of the on-chromatin assembly of the histone-modifying complexes of interest in live cells.For complete details on the use and execution of this protocol, please refer to Cheng et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
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| ISSN: | 2666-1667 |