Institute for Regenerative Medicine and Department of Cell and Developmental Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104-6058, USA.
Centre for Genomic Regulation (CRG), Barcelona Biomedical Research Park, 08003 Barcelona, Spain.
STAR Protoc. 2020 Dec 13;1(3):100223. doi: 10.1016/j.xpro.2020.100223. eCollection 2020 Dec 18.
This protocol provides a two-parameter analysis of single-molecule tracking (SMT) trajectories of Halo-tagged histones in living adherent cell lines and unveils a chromatin mobility landscape composed of five chromatin types, ranging from low to high mobility. When the analysis is applied to Halo-tagged, chromatin-binding proteins, it associates chromatin interaction properties with known functions in a way that previously used SMT parameters did not. For complete information on the use and execution of this protocol, please refer to Lerner et al. (2020).
本方案提供了一种双参数分析方法，用于分析活贴壁细胞系中 Halo 标记组蛋白的单分子追踪（SMT）轨迹，揭示了由五种不同流动性的染色质类型组成的染色质流动性图谱。当将该分析应用于 Halo 标记的染色质结合蛋白时，它将染色质相互作用特性与先前使用的 SMT 参数所没有的已知功能联系起来。有关本方案的使用和执行的完整信息，请参考 Lerner 等人（2020 年）。
