Mesoscale proximity labeling at chromatin to study small molecule mechanism of action [CUTandTag_2]

Proximity labeling traditionally identifies interactomes of a single protein or RNA, though this approach limits mechanistic understanding of biomolecules functioning within complex systems. Here, we demonstrate a strategy for deciphering ligand-induced changes to global biomolecular interactions by enabling proximity labelling at the mesoscale, across an entire cellular system. By inserting nanoscale proximity labelling catalysts throughout chromatin, this system, MesoMap, provided new insights into how HDAC inhibitors regulate gene expression. Furthermore, it revealed that the orphaned drug candidate, SR-1815, regulates disease-linked Syngap1 gene expression through direct inhibition of kinases implicated in both neurological disorders and cancer. Through precise mapping of global chromatin mobility, MesoMap promotes insights into how drug-like chemical probes induce transcriptional dynamics within healthy and disease-associated cellular states. Overall design: pc293T cells, which contained FLAG tagged histone constructs, under went CUT&Tag sequencing to confirm localization of histone constructs thoughout chromatin. N=3 samples were used for each antibody target. The antibodies included FLAG, Histone H3.3, and IgG.