Programmable human histone phosphorylation and gene activation using a CRISPR/Cas9-based chromatin kinase

We report a synthetic chromatin kinase, by fusing nuclease-null CRISPR/Cas9 to a hyperactive, truncated variant of the human MSK1 histone kinase (dCas9-dMSK1). To investigate the role of MSK1 on transcriptional regulation, we explore the different transcriptome between wildtype and MSK1 knockout HEK 293T cell lines by RNA-seq. To test the efficacy of dCas9-dMSK1 at thousands of human promoters in high-throughput and to evaluate if histone phosphorylation could uncover novel mediators of pathological gene expression, we performed a CRISPRa screening assay in A375 cells using the genome wild gRNA library. The enriched gRNAs were measured by Miseq. Examined the transcriptional activation role of native MSK1 and synthetic dCas9-dMSK1 in different cell lines.