Journal: ACS Infectious Diseases

Article Title: A Multiplexed, Target-Based Phenotypic Screening Platform Using CRISPR Interference in Mycobacterium abscessus

doi: 10.1021/acsinfecdis.5c00623

Figure Lengend Snippet: Growth of the 60 hypomorph strains generated in single-plex and multiplex. (A) Strain growth rates measured in single-plex. Growth (%) was calculated based on OD 600 values in liquid medium normalized to growth under uninduced CRISPRi conditions after 4 days (blue). Growth defects were seen in a majority of hypomorphs when CRISPRi was induced with 100 ng/mL anhydrotetracycline (ATc) (red). Ciprofloxacin (CIP) was used as a positive well-killing control (purple). Data are mean values and standard deviations from four technical replicates. (B) Growth assay in multiplex. Strain abundance was measured using read counts as a surrogate after normalization to account for any variation in DNA extraction and PCR amplification between and within plates. All cultures were supplemented with 100 ng/mL ATc as above to induce CRISPRi. CIP was used as a positive well-killing control (red) while DMSO was used as a negative control (blue). Median normalized read counts were obtained from 12 wells per plate. Data represents the mean values and SEM from seventy-two technical replicate plates.

Article Snippet: 1% DMSO (Sigma-Aldrich) was used as the negative untreated control and 25 μg/mL ciprofloxacin hydrochloride (CIP) (MP Biomedicals) was used as the positive control.

Techniques: Generated, Multiplex Assay, Control, Growth Assay, DNA Extraction, Amplification, Negative Control

Journal: Nature Microbiology

Article Title: A host-directed adjuvant sensitizes intracellular bacterial persisters to antibiotics

doi: 10.1038/s41564-025-02124-2

Figure Lengend Snippet: a , b , KL1 boosted Rif activity against clinical M. tuberculosis strains from the globally prevalent lineage 2 ( a ) and lineage 4 ( b ) in macrophages. C.f.u.s were normalized to the untreated control (Gen-only) ( n = 3, two-sided unpaired t -test). c , KL1 enhanced ciprofloxacin (Cip; 5 µg ml −1 ) killing of intracellular S . Typhimurium. Surviving bacteria were normalized to input c.f.u.s ( n = 7, two-sided unpaired t -test). Bars represent mean ± s.e.m. d , Schematic of the Salmonella murine infection model. C57BL/6J mice were infected via oral gavage and intraperitoneally treated with 150 mg kg −1 cefotaxime (CTX) with or without 100 mg kg −1 KL1 (b.i.d.) at 2 dpi for 6 days. Created with BioRender.com . e – h , Tissue samples were collected to enumerate the number of surviving bacteria (c.f.u.s) at 4 dpi ( e ) and 8 dpi ( f ). Co-administration of KL1 and CTX reduced the bacterial burden in Peyer’s patches (PP) ( e , f , h ) but not in mesenteric lymph nodes (MLN) ( e – g ). A group of 5 mice was examined over the time course ( n = 5, two-sided unpaired t -test). Bars represent mean ± s.e.m.

Article Snippet: Cells were washed once with PBS and treated with 5 μg ml −1 ciprofloxacin (MP Biomedicals) in the presence of 10–80 μM KL1 or 0.2% DMSO for 24 h. Cells were washed three times in 500 μl PBS, lysed with 0.2% Triton X-100 at 37 °C for 10 min and plated on LB agar.

Techniques: Activity Assay, Control, Bacteria, Infection