TurboCas: A method for locus-specific labeling of genomic regions and isolating their associated protein interactome.

ABSTRACT: Regulation of gene expression during development and stress response requires the concerted action of transcription factors and chromatin-binding proteins. Because this process is cell-type specific and varies with cellular conditions, mapping of chromatin factors at individual regulatory loci is crucial for understanding cis-regulatory control. Previous methods only characterize static protein binding. We present "TurboCas," a method combining a proximity-labeling (PL) enzyme, miniTurbo, with CRISPR-dCas9 that allows for efficient and site-specific labeling of chromatin factors in mammalian cells. Validating TurboCas at the FOS promoter, we identify proteins recruited upon heat shock, cross-validated via RNA polymerase II and P-TEFb immunoprecipitation. These methodologies reveal canonica

SUBMITTER: Cenik BK 

PROVIDER: S-EPMC12327812 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature