Fig. 1: Development of a CRISPR/Cas9-based histone kinase. | Nature Communications

Fig. 1: Development of a CRISPR/Cas9-based histone kinase.

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

Fig. 1

a Schematics of dCas9, dCas9 fused to WT MSK1 (dCas9-MSK1), dCas9 fused to a hyperactive truncated MSK1 (dCas9-dMSK1), and dCas9 fused to a catalytically inactivated version of dMSK1 (dCas9-ddMSK1). b dCas9 and dCas9-MSK1 fusion variants were transiently transfected into HEK293T cells and autophosphorylation levels were detected (at serine residues 212 and 376 of MSK1) by Western blot 72 h post-transfection. Data are representative of three independent biological experiments. c Purified dCas9, dCas9-MSK1, dCas9-dMSK1, and dCas9-ddMSK1 were incubated with human histone octamers in vitro and resulting levels of histone H3 phosphorylation at serine residues 10 and 28 (H3S10ph and H3S28ph, respectively) were measured after incubation for 1 h. d The relative levels of H3S10ph and H3S28ph were quantified using densitometry from three different in vitro histone kinase assays. Two-sided t-test, *P < 0.05; n = 3 independent experiments in panel (d); error bars, s.e.m.; ns, not significant; kDa, kilodaltons. Source data are available in the Source data file.

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