Fig. 6: Genome-scale screening using dCas9-dMSK1 identifies mediators of BRAF V600E inhibitor resistance. | Nature Communications

Fig. 6: Genome-scale screening using dCas9-dMSK1 identifies mediators of BRAF V600E inhibitor resistance.

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

Fig. 6

a Flow-chart of the genome-scale screening regime using dCas9-dMSK1. Puro; puromycin. b Box plots showing the distribution of gRNA frequencies after lentiviral transduction in DMSO or PLX-4720 treated A375 cells. In all box plots, the green horizontal line represents the median and the upper and lower bounds of each box correspond to the 25th and 75th percentiles of each sample, respectively. The upper and lower whiskers extend to the maximum and minimum values, respectively, within 1.5-fold of the interquartile range of the box bounds. Outliers are plotted as green dots. n = 2 independent experiments. c Volcano plot displaying gRNA counts in PLX-4720 versus DMSO treated A375 cells. Data were analyzed using robust rank aggregation (RRA) with a cutoff of P value <0.01. Blue circles indicate depleted gRNAs, red circles indicate enriched gRNAs, and gray circles indicate gRNAs with no change. d mRNA levels of the top 10 most enriched genes in A375 cells stably expressing dCas9 or dCas9-dMSK1 when targeted with corresponding gRNAs identified from the screen. Two-sided t-test, *P < 0.05; n = 3 independent experiments for panel (d); error bars, s.e.m. Source data are available in the Source data file.

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