Protocol for detecting chromatin dynamics and screening chromatin relaxer by FRAP assay

STAR Protocols

Keshi Chen¹  Juntao Qi¹  Hao Wu²  Wei Li²  Xingguo Liu³  Yanshuang Zhou³  Qi Long³ 

[1] Bioland laboratory (Guangzhou Regenerative Medicine and Health Guangdong Laboratory), Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, CUHK-GIBH Joint Research Laboratory on Stem Cells and Regenerative Medicine, Institute for Stem Cell and Regeneration, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China;University of Chinese Academy of Sciences, Beijing 100049, China;CAS Key Laboratory of Regenerative Biology, Joint School of Life Sciences, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou Medical University, Guangzhou 510530, China;

关键词: Cell Biology;    Cell-based Assays;    Microscopy;    Molecular Biology;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: We describe a fluorescence recovery after photobleaching (FRAP) protocol for assessing the dynamics of heterochromatin/euchromatin and identifying chromatin relaxers for cell fate transition. Here, we developed a system to track heterochromatin foci with HP1α-cherry and performed FRAP assay of H1-GFP to analyze the dynamics of heterochromatin and euchromatin during somatic cell reprogramming. This protocol is used to screen factors that impact chromatin structure, which could also be used to identify chromatin relaxers and repressors in various cell fate transitions.For complete details on the use and execution of this protocol, please refer to Chen et al. (2016) and Chen et al. (2020).

【 授权许可】

Unknown