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Fig. 2 | Genome Biology

Fig. 2

From: The epigenome: the next substrate for engineering

Fig. 2

Interrogating the contextual and combinatorial principles of epigenome regulation. a A method for the parallel monitoring of the transcriptional activities of thousands of randomly integrated, barcoded reporters was used to study chromatin position effects across the genome. b Synthetic chromatin regulators (synCRs), composed of fusions of programmable zinc fingers (ZFs) and subunit proteins derived from diverse chromatin-regulating complexes, were used to study and program transcriptional outputs produced by individual and combinations of CRs at integrated reporters. GFP green fluorescent protein. c CRISPR/dCas9 can be exploited for high-throughput functional assays of epigenetic regulators thanks to its experimental tractability for combinatorial and multiplexed recruitment. Scaffolding multiple RNA-hairpin motifs to a guide RNA (gRNA) allows multivalent recruitment of chromatin regulators (CRs). Scaffolding different RNA motifs to gRNA allows heterologous recruitment of CRs. The same CR can be simultaneously recruited to multiple loci by using different gRNAs specific to each gRNA locus

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