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. Author manuscript; available in PMC: 2017 Jun 2.
Published in final edited form as: Cell. 2016 May 26;165(6):1493–1506. doi: 10.1016/j.cell.2016.05.003

Figure 2. CRISPRi Knockdowns of Essential Genes Enable Discovery of Direct Antibiotic Targets.

Figure 2

A) Relative fitness of CRISPRi essential gene knockdown strains (n = 289) with basal dcas9 expression (no xylose induction) grown on plates containing MAC-0170636, as determined by the ratio of normalized colony sizes on LB plates + DMSO versus LB + MAC-0170636.

B) Minimal inhibitory concentration (MIC) assay for strains over- or under-expressing uppS grown in liquid medium containing MAC-0170636. The MICs of these strains were (in μg/ml): sgRNAuppS + 0.05% xyl (∼0.012), sgRNAuppS (∼0.195), WT (∼0.78), and uppS overexpression (∼3.125). The values plotted are the means of at least three measurements, and error bars are ±1 standard deviation.

C) Concentration-dependent inhibition of purified B. subtilis UppS by MAC-0170636. Each point is an individual measurement.

D) MIC assay for strains expressing B. subtilis uppS (BsΔuppS/amyE∷Pspank-uppSBs) or S. aureus uppS (BsΔuppS/amyE∷Pspank-uppSSa) grown in liquid LB + MAC-0170636.