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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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725000036 |
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| author | Shanli Guan Jiajia Tang Cuixia Di Bo Cheng |
| author_facet | Shanli Guan Jiajia Tang Cuixia Di Bo Cheng |
| author_sort | Shanli Guan |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-6335ce2b258d4761a8a6ed0c33f92349 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-6335ce2b258d4761a8a6ed0c33f923492025-01-29T05:02:00ZengElsevierSTAR Protocols2666-16672025-03-0161103597Protocol for visualizing the chromatin assembly properties of epigenetic protein complexes via an HTM module-mediated artificial tethering systemShanli Guan0Jiajia Tang1Cuixia Di2Bo Cheng3School of Life Sciences, Lanzhou University, Lanzhou 730000, Gansu, P.R. ChinaSchool of Life Sciences, Lanzhou University, Lanzhou 730000, Gansu, P.R. ChinaBio-Medical Research Center, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000, Gansu, P.R. ChinaSchool of Life Sciences, Lanzhou University, Lanzhou 730000, Gansu, P.R. China; Key Laboratory of Cell Activities and Stress Adaptations, Ministry of Education, Lanzhou University, Lanzhou 730000, Gansu, P.R. China; Corresponding authorSummary: 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.http://www.sciencedirect.com/science/article/pii/S2666166725000036cell biologymolecular biologybiotechnology and bioengineering |
| spellingShingle | Shanli Guan Jiajia Tang Cuixia Di Bo Cheng Protocol for visualizing the chromatin assembly properties of epigenetic protein complexes via an HTM module-mediated artificial tethering system STAR Protocols cell biology molecular biology biotechnology and bioengineering |
| title | Protocol for visualizing the chromatin assembly properties of epigenetic protein complexes via an HTM module-mediated artificial tethering system |
| title_full | Protocol for visualizing the chromatin assembly properties of epigenetic protein complexes via an HTM module-mediated artificial tethering system |
| title_fullStr | Protocol for visualizing the chromatin assembly properties of epigenetic protein complexes via an HTM module-mediated artificial tethering system |
| title_full_unstemmed | Protocol for visualizing the chromatin assembly properties of epigenetic protein complexes via an HTM module-mediated artificial tethering system |
| title_short | Protocol for visualizing the chromatin assembly properties of epigenetic protein complexes via an HTM module-mediated artificial tethering system |
| title_sort | protocol for visualizing the chromatin assembly properties of epigenetic protein complexes via an htm module mediated artificial tethering system |
| topic | cell biology molecular biology biotechnology and bioengineering |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725000036 |
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