FIG 2.
Genome-wide screening for DNT receptors by the CRISPR-Cas9 system. (A) Procedures for genome-wide screening for the DNT receptor(s). (B) Genes identified after screening. The y and x axes represent the numbers of unique sgRNAs and sgRNA sequence reads for each identified gene, respectively. (C) Sensitivities of the candidate-gene knockout clones of MC3T3-El to DNT-DTA. Seven knockout clones were selected for each gene and treated with DNT-DTA, and the rate of cell death was evaluated. Plotted data represent means ± SEM (n = 3). *, P < 0.0001 (compared to MC3T3-E1 parental cells [E1]). (D) DNT-induced deamidation of Rho in the knockout clones of MC3T3-E1. Two of the seven clones shown in panel C for each gene were further selected and treated with DNT. The numbers of selected clones are shown after the gene names.