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

• Main Authors: Qi Long, Juntao Qi, Wei Li, Yanshuang Zhou, Keshi Chen, Hao Wu, Xingguo Liu
• Format: Article
• Language: English
• Published: Elsevier 2021-09-01
• Series: STAR Protocols
• Subjects: Cell Biology; Cell-based Assays; Microscopy; Molecular Biology
• Online Access: http://www.sciencedirect.com/science/article/pii/S2666166721004135

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).