Figure 10. Phenoprinting can inform antimycobacterial MOA.
Cells were exposed to varying supra-MIC concentrations (1X, 2X, 4X MIC) of the selected antimycobacterial compounds for 18 hr, imaged, and analyzed using the pipeline developed for CRISPRi-imaging. The resulting profiles were visualized in CRISPRi-generated UMAP space (A), and the morphological nearest-neighbor identified (B). Known targets were simultaneously visualized for comparative purposes. BDQ, bedaquiline; EMB, ethambutol; INH, isoniazid; RIF, rifampicin; MOXI, moxifloxacin.
Figure 10—figure supplement 1. Phenoprinting can inform antimicrobial MOA.
Cells were exposed to varying supra-MIC concentrations (1X, 2X, 4X MIC) of the selected antimycobacterial compounds for 18 hr, imaged, and analyzed using the same pipeline developed for CRISPRi-imaging. The resulting profiles were visualized in CRISPRi-generated UMAP space. CIP, ciprofloxacin; DCS, D-cycloserine; GRS, griselimycin; KAN, kanamycin; LEVO, levofloxacin; LNZ, linezolid; NOVO, novobiocin; NRG, nargenicin; OFX, ofloxacin; STR, streptomycin; TMP, trimethoprim; VAN, vancomycin.
Figure 10—figure supplement 2. Phenoprinting can inform antimicrobial MOA.
Cells were exposed to varying supra-MIC concentrations (1X, 2X, 4X MIC) of the selected antimycobacterial compounds for 18 hr, imaged, and analyzed using the same pipeline developed for CRISPRi-imaging. The resulting profiles were visualized in CRISPRi-generated UMAP space. For each compound, representative cells, known targets and the morphological nearest neighbor are presented. CIP, ciprofloxacin, DCS, D-cycloserine; GRS, griselimycin; KAN, kanamycin, LEVO, levofloxacin; LNZ, linezolid. NOVO, novobiocin; NRG, nargenicin; OFX, ofloxacin; STR, streptomycin; TMP, trimethoprim; VAN, vancomycin. *The major spontaneous resistance gene, if available. For kanamycin, rrs is not present in the library. Limited compound availability restricted nargenicin assays to 1X MIC.
Figure 10—figure supplement 3. Phenoprinting can inform antimicrobial MOA.
