A modular dCas9-based recruitment platform for combinatorial epigenome editing

Chromatin immunoprecipitation sequencing (ChIP-seq) for histone modifications H3K4me3, H3K9me3 and H3K27me3 in HEK293T cells stably expressing dCas9-SSSavi and 6x sgRNA, and transfected with either DNMT3A+KRAB+EZH2 (D3A-K-E) or aGCN4-mCherry (non-catalytic control) for 72h, followed by flow cytometric enrichment of 200,000 cells positive for GFP (containing SSSavi components).
WGBS was performed on HSB6G cells transfected with either D3A-K-E or 𝛼GCN4-mCherry for 72h, followed by flow cytometric enrichment of 100,000 cells positive for GFP (containing SSSavi components). Genomic DNA was extracted and processed as detailed in the protocols section.